American Chemical Society - Division of Carbohydrate Chemistry ...
American Chemical Society - Division of Carbohydrate Chemistry ...
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<strong>American</strong> <strong>Chemical</strong> <strong>Society</strong><br />
<strong>Division</strong> <strong>of</strong> <strong>Carbohydrate</strong> <strong>Chemistry</strong><br />
240th ACS National Meeting, Boston, MA, August 22-26, 2010<br />
T. Lowary, Program Chair<br />
SUNDAY MORNING<br />
Wolfrom-Isbell-New Investigator Award Symposium<br />
G. Eggleston, Organizer; T. Lowary, Organizer; T. Lowary, Presiding Papers 1-3<br />
SUNDAY AFTERNOON<br />
Wolfrom-Isbell-New Investigator Award Symposium<br />
G. Eggleston, Organizer; T. Lowary, Organizer; T. Lowary, Presiding Papers 4-9<br />
MONDAY MORNING<br />
Synthetic Oligosaccharides and Glycoconjugates for Preventing and<br />
Combating Disease<br />
L. Wang, Organizer; G. Boons, Organizer; G. Boons, Presiding Papers 10-16<br />
MONDAY AFTERNOON<br />
Synthetic Oligosaccharides and Glycoconjugates for Preventing and<br />
Combating Disease<br />
G. Boons, Organizer; L. Wang, Organizer; L. Wang, Presiding Papers 17-22<br />
MONDAY EVENING<br />
Sci-Mix<br />
T. Lowary, Organizer Papers 85, 89, 86, 92, 87, 94, 93, 110, 108, 115, 96, 113,<br />
83, 98, 82, 97, 100, 81, 102, 88, 103, 101, 104, 111, 106, 105, 107<br />
TUESDAY MORNING<br />
Petite and Sweet: Glyconanotechnology as a Bridge to New Medicines<br />
J. Barchi, Organizer; X. Huang, Organizer; J. Barchi, Presiding Papers 23-27<br />
Oligonucleotide Therapeutics
M. Manoharan, Organizer; M. Manoharan, Presiding; R. Iyer, Presiding Papers<br />
28-38<br />
TUESDAY AFTERNOON<br />
Petite and Sweet: Glyconanotechnology as a Bridge to New Medicines<br />
J. Barchi, Organizer; X. Huang, Organizer; J. Barchi, Presiding Papers 39-44<br />
Oligonucleotide Therapeutics<br />
M. Manoharan, Organizer; M. Manoharan, Presiding; S. Srivastava, Presiding<br />
Papers 45-54<br />
WEDNESDAY MORNING<br />
Recognition <strong>of</strong> DNA: Recent Advances<br />
D. Arya, Organizer; D. Arya, Presiding Papers 55-59<br />
General Papers<br />
T. Lowary, Organizer; E. Ci<strong>of</strong>fi, Presiding Papers 60-66<br />
WEDNESDAY AFTERNOON<br />
General Papers<br />
T. Lowary, Organizer; S. Graham, Presiding Papers 67-74<br />
Recognition <strong>of</strong> DNA: Recent Advances<br />
D. Arya, Organizer; D. Arya, Presiding Papers 75-79<br />
WEDNESDAY EVENING<br />
General Posters<br />
T. Lowary, Organizer Papers 80-115<br />
THURSDAY MORNING<br />
General Papers<br />
T. Lowary, Organizer; C. Marzabadi, Presiding Papers 116-124<br />
THURSDAY AFTERNOON
General Papers<br />
T. Lowary, Organizer; R. Narain, Presiding Papers 125-135
CARB: <strong>Division</strong> <strong>of</strong> <strong>Carbohydrate</strong> <strong>Chemistry</strong> S M T W T<br />
General Papers D D<br />
General Posters E<br />
Oligonucleotide Therapeutics** D<br />
Petite and Sweet: Glyconanotechnology as a Bridge to New Medicines D<br />
Recognition <strong>of</strong> DNA: Recent Advances P<br />
Recognition <strong>of</strong> DNA: Recent Advances** A<br />
Sci-Mix E<br />
Synthetic Oligosaccharides and Glycoconjugates for Preventing and<br />
Combating Disease<br />
Wolfrom-Isbell-New Investigator Award Symposium D<br />
Legend<br />
A = AM; P = PM; D = AM/PM; E = EVE;<br />
AE = AM/EVE; DE = AM/PM/EVE; PE = PM/EVE;<br />
*Cosponsored symposium with primary organizer shown in parenthesis; located with primary organizer.<br />
**Primary organizer <strong>of</strong> cosponsored symposium.<br />
D
CARB Todd Lowary Sunday, August 22, 2010<br />
Oral<br />
Wolfrom-Isbell-New Investigator Award Symposium - AM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Gillian Eggleston, Todd Lowary<br />
Presiders: Todd Lowary<br />
Duration: 9:15 am - 11:20 am<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
9:15 am Introductory Remarks<br />
9:20 am 1 Developing an enzymatic approach to synthesize heparan sulfates<br />
Jian Liu<br />
10:00 am 2 Creating and implementing breakthrough medical technology<br />
Dr Robert S Langer<br />
10:40 am 3 Advances in heparin analysis and synthesis<br />
Pr<strong>of</strong>. Robert J. Linhardt PhD
CARB Todd Lowary Sunday, August 22, 2010<br />
Wolfrom-Isbell-New Investigator Award Symposium - PM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Gillian Eggleston, Todd Lowary<br />
Presiders: Todd Lowary<br />
Duration: 1:00 pm - 4:45 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
1:00 pm 4 Development <strong>of</strong> a fully synthetic three-component carbohydrate-based cancer<br />
vaccine<br />
pr<strong>of</strong>. Geert-Jan Boons PhD<br />
1:35 pm 5 Chemoenzymatic synthesis and antibody recognition <strong>of</strong> HIV-1 V3 glycopeptides<br />
Pr<strong>of</strong>. Lai-Xi Wang<br />
2:10 pm 6 Entirely carbohydrate vaccine constructs and their application in probing<br />
glycoimmunology<br />
Pr<strong>of</strong>. Peter R Andreana PhD<br />
2:45 pm Intermission<br />
3:00 pm 7 <strong>Carbohydrate</strong> polymer assembly: How do mycobacteria do it?<br />
Laura L. Kiessling<br />
3:35 pm 8 Lipopolysaccharide transport and assembly in Escherichia coli<br />
Pr<strong>of</strong>. Daniel Kahne<br />
4:10 pm 9 Arrays and automated oligosaccharide synthesis<br />
Pr<strong>of</strong>. Nicola L. B. Pohl
CARB Todd Lowary Monday, August 23, 2010<br />
Oral<br />
Synthetic Oligosaccharides and Glycoconjugates for Preventing and Combating Disease - AM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Lai-Xi Wang, GJ Boons<br />
Presiders: GJ Boons<br />
Duration: 8:00 am - 12:00 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
8:00 am Introductory Remarks<br />
8:05 am 10 Bioactive N-linked glycopeptides and glycoprotein conjugates<br />
Thomas J. Tolbert PhD<br />
8:25 am 11 Efficient chemoenzymatic synthesis <strong>of</strong> sialyl Lewis antigens using<br />
sialyltransferase mutants<br />
Pr<strong>of</strong>essor/Ph.D. Xi Chen PhD, Go Sugiarto, Kam Lau, Lei Zhang, Dr. Shengshu<br />
Huang Ph.D.<br />
9:00 am 12 Synthesis <strong>of</strong> saponin vaccine adjuvants for immunotherapy<br />
Pr<strong>of</strong>. David Y Gin PhD<br />
9:35 am 13 Glycosylation engineering <strong>of</strong> human IgG-Fc for functional studies<br />
Pr<strong>of</strong>. Lai-Xi Wang
10:10 am Intermission<br />
10:30 am 14 Chemoenzymatic synthesis <strong>of</strong> GPI-anchored proteins and glycoproteins<br />
Dr. Xueqing Guo, Dr. Zhimeng Wu, Pr<strong>of</strong>. Zhongwu Guo<br />
10:50 am 15 Cancer relevant epitopes uncovered by synthetic mucin glycopeptides<br />
Pr<strong>of</strong>. Shin-Ichiro Nishimura PhD<br />
11:25 am 16 <strong>Chemical</strong> biology <strong>of</strong> O-GlcNAc processing enzymes<br />
Pr<strong>of</strong>. David J Vocadlo, Dr. Tracey M Gloster, Wesley F Zandberg, Julia E<br />
Heinonen, David Shen, Thomas Clark, Dr. Carlos Martinez-Fleites, Yuan He,<br />
Pr<strong>of</strong>. Gideon J Davies<br />
CARB Todd Lowary Monday, August 23, 2010<br />
Oral<br />
Synthetic Oligosaccharides and Glycoconjugates for Preventing and Combating Disease - PM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: GJ Boons, Lai-Xi Wang<br />
Presiders: Lai-Xi Wang<br />
Duration: 1:30 pm - 5:05 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
1:30 pm 17 Synthesis <strong>of</strong> a single molecule L-rhamnose-containing three component vaccine<br />
and evaluation <strong>of</strong> antigenicity in the presence <strong>of</strong> anti L-rhamnose antibodies
1:50 pm 18<br />
M.S. Sourav Sarkar, M.S. Rommel S Talan, Mr. Steven A Lombardo, Pr<strong>of</strong>.<br />
Katherine A Wall PhD, Pr<strong>of</strong>. Steven J Sucheck PhD<br />
Design <strong>of</strong> glycoprotein vaccines against influenza and other viral diseases<br />
Pr<strong>of</strong>. Chi-Huey Wong PhD<br />
2:25 pm 19 Modular synthesis <strong>of</strong> heparan sulfate oligosaccharides for array development<br />
Pr<strong>of</strong>. Geert-Jan Boons<br />
3:00 pm Intermission<br />
3:15 pm 20 New targets for antibiotics<br />
Pr<strong>of</strong>essor Suzanne Walker<br />
3:50 pm 21 Challenges and opportunities in natural product glycosylation<br />
Jon S Thorson<br />
4:25 pm 22 Glycomimetic and non-carbohydrate inhibitors as probes <strong>of</strong> the lectin DC-SIGN<br />
Pr<strong>of</strong>essor Laura L Kiessling<br />
5:00 pm Concluding Remarks
CARB Todd Lowary Monday, August 23, 2010<br />
Sci-Mix<br />
Sci-Mix - EVE Session<br />
Location: Boston Convention & Exposition Center<br />
Room: Hall C<br />
Organizers: Todd Lowary<br />
Duration: 8:00 pm - 10:00 pm<br />
Pub<br />
Presentation Title<br />
#<br />
81 Expression and characterization <strong>of</strong> enzymes for bioenzymatic synthesis <strong>of</strong> heparin<br />
Priscilla Paul, Wenjing Zhao, Pr<strong>of</strong>. Robert Linhardt, Pr<strong>of</strong>. Jonathan Dordick, Pr<strong>of</strong>. Jian Liu<br />
82 Catalytic conversion <strong>of</strong> biomass-derived carbohydrates to useful chemicals in one step<br />
Dr. Weiran Yang, Pr<strong>of</strong>. Ayusman Sen<br />
83 Behaviour <strong>of</strong> polysaccharide aggregates in asymmetricalfield-flow fractionation and sizeexclusion<br />
chromatography<br />
Leena Pitkänen, Maija Tenkanen, Päivi Tuomainen<br />
85 Synthesis and structural optimization <strong>of</strong> antifungal kanamycin B analogs<br />
Marina Y FOSSO, Yukie KAWASAKI, Sanjib SHRESTHA, Jon TAKEMOTO, Tom C.-<br />
W. CHANG<br />
86 E. coli K5 heparosan production for bioengineered heparin<br />
Zhenyu Wang, Mellisa Ly, Fuming Zhang, Zhenqing Zhang, Jonathan S. Dordick, Robert J.<br />
Linhardt
87 Stochastic simulation <strong>of</strong> lectin microarrays with nanosensor transducers: Potential platforms<br />
for optimal, high-throughput screening and pr<strong>of</strong>iling <strong>of</strong> glycoproteins<br />
Mr. Nigel F Reuel, Dr. Jin-Ho Ahn PhD, Dr. Michael S. Strano PhD<br />
88 Evaluation <strong>of</strong> different thioesters for glycocluster synthesis applying native chemical<br />
ligation<br />
Johannes W. Wehner, Pr<strong>of</strong>. Dr. Thisbe K. Lindhorst<br />
89 NMR for structure elucidation <strong>of</strong> commercially available heparin polysaccharides<br />
Kemal Solakyildirim, Scott A. McCallum, Robert J. Linhardt<br />
92 Study on therelative reactivity <strong>of</strong> glycosyl acceptors in the glycosylations <strong>of</strong>2-Azido-2deoxy-galactosides<br />
Jane Kalikanda, Dr. Zhitao Li<br />
93 Synthesis <strong>of</strong> tailored glycoconjugates for the precise detection <strong>of</strong> pathogens<br />
Ashish A. Kulkarni, Dr. Suri S. Iyer<br />
94 Structure-activity studies <strong>of</strong> synthetic glycophosphatidylinositol anchored proteins<br />
Dr. Carl V Christianson, Pr<strong>of</strong>. Peter H Seeberger<br />
96 Synthesis <strong>of</strong> a fluorous-tagged disaccharide for the enzymatic preparation <strong>of</strong> heparin<br />
oligosaccharides<br />
Sayaka Masuko, Smritilekha Bera, Robert J. Linhardt<br />
97 Preventing the transmission <strong>of</strong> Plasmodium falciparum through the inhibition <strong>of</strong> malaria<br />
protein binding to placental chondroitin sulfate A<br />
Julie M Beaudet, Leandra J Mansur, Bo Yang, Fuming Zhang, Robert J Linhardt<br />
98 Real-time assessment <strong>of</strong> the morphological change <strong>of</strong> cellulose in response to enzymatic<br />
treatment<br />
Chi Nguyen<br />
100 Immobilization <strong>of</strong> enzymes relevant to bioengineered heparin synthesis<br />
Eric R. Sterner, Dr. Robert J. Linhardt, Dr. Jonathan S. Dordick, Dr. Jian Liu, Dr. Fuming<br />
Zhang, Wenjing Zhao, Priscilla Paul, Jeff Martin<br />
101 Analyses <strong>of</strong> anti Tn-antigen MLS128 monoclonal antibody binding to two or three
consecutive Tn-antigen clusters by surface plasmon resonance (SPR) and NMR<br />
Ayano Takasaki-Matsumoto Ph.D., Shinya Hanashima Ph.D., Ami Aoki, Yoshiki<br />
Yamaguchi Ph.D., Reiko Sato, Hiroko Kawakami Ph.D., Mamoru Mizuno Ph.D., Pr<strong>of</strong>.<br />
Yoko Fujita-Yamaguchi Ph.D.<br />
102 Modular glycoconjugate tool set for assembly and presentation <strong>of</strong> multivalent carbohydrate<br />
ligands on surfaces<br />
Irene Abia, Brian Sanders, Michael D. Best, David C. Baker<br />
103 Analysis<strong>of</strong> the absorption <strong>of</strong> low molecular weight heparin in human umbilical cord tissue<br />
as a model for the prevention <strong>of</strong> cancer metastasis<br />
Amanda M. Weyers, Thangriala Sudha, Bo Yang, Boyangzi Li, Majde Takieddin, Fuming<br />
Zhang, Shaker A. Mousa, Robert J. Linhardt<br />
104 Synthesis and biological evaluation <strong>of</strong> a Gal(α1-2)GalCer analog<br />
Yanke Liang, Pr<strong>of</strong>. Amy R Howell PhD<br />
105 Thiol-click chemistry approach to glycomimetics: Novel stereoselective synthesis <strong>of</strong> (1-3)-<br />
S-thiodisaccharides<br />
Pr<strong>of</strong>. Zbigniew J. Witczak, Irena Bak-Sypien<br />
106 High-throughput glycoarray for monitoring immune responses to a cancer vaccine<br />
Dr. Christopher Campbell M.D., Ph.D., Dr. Yalong Zhang Ph.D., Dr. Olaf Ludek Ph.D.,<br />
Mr. David Farnsworth, Dr. Jeffrey Gildersleeve Ph.D.<br />
107 NMR spectroscopic studies <strong>of</strong> APF: A small glycopeptide possessing potent<br />
antiproliferative activity<br />
Dr Kristie M Adams PhD, Dr Piotr Kaczmarek PhD, Dr Susan K Keay MD, PhD, Dr<br />
Joseph J Barchi, Jr PhD<br />
108 Structural and quantitative analysis <strong>of</strong> disaccharides using CE with LIF detection<br />
Yuqing Chang, Tatiana Laremore, Fuming Zhang, Robert J Linhardt<br />
110 Carbon nanotubes and chitosan as possible scaffolds for bone tissue regeneration<br />
Julia Stone, Yetunde Olusanya, Whitney Jones, Pasakorn Traisawatwong, Melisa Stewart,<br />
Cordella Kelly-Brown, Laura Carson Ph.D., Aderemi Oki Ph.D., E. Gloria C. Regisford<br />
Ph.D.
111 Biosynthesis <strong>of</strong> heparin by metabolic engineering <strong>of</strong> Chinese hamster ovary cells<br />
Leyla Gasimli, Jongyoun Baik PhD, Dr. Susan Sharfstein PhD, Pr<strong>of</strong>. Robert J. Linhardt<br />
PhD<br />
113 Design, synthesis, and characterization <strong>of</strong> sulfated N-aryl aminoglycosides<br />
Amanda M. Fenner, Robert J. Kerns PhD<br />
115 On-line micr<strong>of</strong>low high-performance liquid chromatography with nano-electrospray<br />
ionization mass spectrometry for heparan sulfate disaccharide analysis<br />
Dr. Bo Yang, Kemal Solakyildirim, Jeffrey G. Martin, Tatiana Laremore, Pr<strong>of</strong>. Robert J.<br />
Linhardt
CARB Todd Lowary Tuesday, August 24, 2010<br />
Oral<br />
Petite and Sweet: Glyconanotechnology as a Bridge to New Medicines - AM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Joseph Barchi, Xuefei Huang<br />
Presiders: Joseph Barchi<br />
Duration: 8:30 am - 11:35 am<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
8:30 am Introductory Remarks<br />
8:35 am 23 Toxicity and functional assessment using polysaccharide-based magnetic iron<br />
oxide nanoparticles for cell labeling in vivo and in vitro<br />
Yoshitaka Miyamoto PhD, Yumie Koshidaka, Hiroaki Saito, Yukimasa Kagami,<br />
Katsutoshi Murase, Noritada Kaji, Hiroshi Yukawa, Hir<strong>of</strong>umi Noguchi MD, PhD,<br />
Hisashi Iwata MD, Yoshinobu Baba PhD, Shuji Hayashi MD, PhD<br />
9:05 am 24 Glycodendrimers for the mediation <strong>of</strong> cancer cellular aggregation processes<br />
Dr. Mark L. Wolfenden, Julie J Sprenger, Michael P Capp, Dr. Pratima Nangia-<br />
Makker, Dr. Avraham Raz, Dr. Mary J Cloninger<br />
9:35 am 25 Glyconanoparticles: Multibi<strong>of</strong>unctional nanomaterials for biomedical applications<br />
Pr<strong>of</strong>. Soledad Penades<br />
10:05 am Intermission
10:35 am 26 New glycopeptide-based nanoparticle constructions for anticancer therapy<br />
Dr. Joseph John Barchi Jr, Dr. Ray Brinas, Dr. Andreas Sundgren, Dr. Padmini<br />
Sahoo, Ms. Amy Houghten, Ms Susan Morey, Mr Michael Sanford, Dr. Howard<br />
Young<br />
11:05 am 27 Functionalized catanionic surfactant vesicles: A new approach to carbohydrate<br />
vaccines<br />
Juhee Park, Lenea Rader, Glen Thomas, Douglas English, Lindsey Zimmerman,<br />
Daniel C. Stein, Philip DeShong<br />
CARB Todd Lowary Tuesday, August 24, 2010<br />
Oral<br />
Petite and Sweet: Glyconanotechnology as a Bridge to New Medicines - PM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Joseph Barchi, Xuefei Huang<br />
Presiders: Joseph Barchi<br />
Duration: 1:30 pm - 5:05 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
1:30 pm 39 Sugar and proteins: Applications <strong>of</strong> bioconjugates
2:00 pm 40<br />
Pr<strong>of</strong>essor Benjamin G. Davis<br />
Sialic acid recognition: Particles, chips, and cells<br />
Pr<strong>of</strong>. Robert A. Field<br />
2:30 pm 41 Functionalized nanoparticle for protein detection<br />
Pr<strong>of</strong>. Chun-Cheng Lin<br />
3:00 pm Intermission<br />
3:30 pm 42 Nanotechnology-based tools for the glycomics revolution: Silicon nanophotonic<br />
carbohydrate biosensors<br />
Pr<strong>of</strong>. Daniel M Ratner PhD<br />
4:00 pm 43 Targeted glyco-magnetic nanoprobes for detection and molecular imaging <strong>of</strong><br />
atherosclerosis<br />
Kheireddine El-Boubbou PhD, Dr. Medha Kamat PhD, Pr<strong>of</strong>. David Zhu PhD,<br />
Ruiping Huang, Dr. George Abela MD, Pr<strong>of</strong>. Xuefei Huang PhD<br />
4:30 pm 44 <strong>Carbohydrate</strong>s on nanoparticles, surfaces and polymers: From basics to<br />
diagnostics applications<br />
Pr<strong>of</strong>. Dr. Peter H. Seeberger<br />
5:00 pm Concluding Remarks
CARB Todd Lowary Tuesday, August 24, 2010<br />
Oral<br />
Oligonucleotide Therapeutics - AM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 252A<br />
Cosponsored<br />
by:<br />
ANYL, BIOL, MEDI, ORGN<br />
Organizers: Muthiah Manoharan<br />
Presiders: Muthiah Manoharan, Radhakrishnan Iyer<br />
Duration: 8:40 am - 11:45 am<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
8:40 am Introductory Remarks<br />
8:45 am 28 Novel synthesis <strong>of</strong> 2'-deoxynucleoside 5'-triphosphates without nucleoside<br />
protection and DNA polymerase recognition<br />
Julianne M. Caton-Williams PhD, Matthew R. Smith, Zhen Huang PhD<br />
9:00 am 29 Activation <strong>of</strong> retinoic acidinducible gene (RIG-I) by short oligonucleotides<br />
Radhakrishnan Iyer, John Coughlin, Seetharamaiyer Padmanabhan, Brent<br />
Korba, Sua Myong<br />
9:15 am 30 Solid-phase synthesis <strong>of</strong> 5'-di- and tri-phosphates andtheir modified analogs <strong>of</strong><br />
DNA, RNA and chemically modified oligonucleotides<br />
Ivan Zlatev, Thomas Lavergne, Sudhakar Takkellapati, Rajenda Pandey,
9:30 am 31<br />
Yupeng Fan, Marija Prhavc, Kathy Mills, G. Rajeev Kallanthottathil, Françoise<br />
Debart, Jean-Jacques Vasseur, François Morvana, Muthiah Manoharan<br />
Drug discovery harnessing RNA interference<br />
Muthiah Manoharan<br />
9:45 am 32 Parallel high throughput synthesis <strong>of</strong> chemically modified 21-27mersiRNA<br />
sequences<br />
Jason Costigan, Satya Kuchimanchi, Sarfraz Shaikh, Keri Dufault, Jack de<br />
Groot, Rachel Meyers<br />
10:00 am 33 New developments in the synthesis <strong>of</strong> oligoribonucleotides: Use <strong>of</strong><br />
dimer/trimer blocks in combination with an “ionic tag” soluble support<br />
Matthew R Hassler, Dr. Nandyala Mallikarjuna Reddy, Dr. Tak-Hang Chan,<br />
Dr. Masad J Damha<br />
10:15 am Break<br />
10:30 am 34 Advanced process <strong>of</strong> RNA synthesis<br />
Dr. Anuj Mohan<br />
10:45 am 35 Novel method for the confirmation <strong>of</strong> siRNA sequence byLC-MS/MS<br />
Gary Lavine, Matthias Kretschmer, James McArdle, Satya Kuchimanchi,<br />
Veeragu Murugaiah, Muthiah Manoharan<br />
11:00 am 36 Evaluation <strong>of</strong> Canonical vs.Dicer-substrate siRNAs in vitro and in vivo<br />
Don Foster, Sayda Elbashir, Satya Kuchimanchi, Greg Hinkle, William<br />
Cantley, Rick Duncan, Geff Cole, Chris Sherill, Kathy Mills, Mara Broberg,<br />
Jeff Rollins, Klaus Charisse, Muthiah Manoharan<br />
11:15 am 37 Modulation <strong>of</strong> thermal stability can enhance the potency <strong>of</strong> siRNA<br />
Haripriya Addepalli, Meena Meena, Chang G. Peng, Gang Wang, Yupeng Fan,<br />
Klaus Charisse, K. Narayanannair Jayaprakash, G. Rajeev Kallanthottathil,<br />
Rajendra Pandey, Gary Lavine, Ligang Zhang, Kerstin Jahn-H<strong>of</strong>mann, Philipp<br />
Hadwiger, Muthiah Manoharan, Martni Maier<br />
11:30 am 38 <strong>Carbohydrate</strong> conjugation tosiRNA for tissue-specific delivery
G. Rajeev Kallanthottathil, K. Narayanannair Jayaprakash, Maria Frank-<br />
Kamenetsky, Gang Wang, Tianlei Lei, Mariano Severgnini, William Querbes,<br />
Jim Butler, Alfica Sehgal, Tomoko Nakayama, Klaus Charrise, Martin Maier,<br />
Kevin Fitzgerald, Muthiah Manoharan<br />
CARB Todd Lowary Tuesday, August 24, 2010<br />
Oral<br />
Oligonucleotide Therapeutics - PM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 252A<br />
Cosponsored<br />
by:<br />
ANYL, BIOL, MEDI, ORGN<br />
Organizers: Muthiah Manoharan<br />
Presiders: Muthiah Manoharan, Suresh Srivastava<br />
Duration: 2:00 pm - 4:50 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
2:00 pm 45 Efficient synthesis <strong>of</strong> siRNA-folic acid conjugates<br />
Rajendra Pandey, Muthusamy Jayaraman, Anna Borodovsky, David Butler,<br />
Shigeo Matsuda, Gang Wang, Martin Maier, Bo Peng, Marjorie Solomon,<br />
Sergey Shulga-Morskoy, Klaus Charrise, David Bumcrot, Kristina Yucius,<br />
Victor Kotelianski, G. Rajeev Kallanthottathil, Muthiah Manoharan
2:15 pm 46 Combining 2'-TBDMS and 2'-ALE chemistry for on-column site specific<br />
modifications <strong>of</strong> RNA<br />
Dr. Jeremy G Lackey PhD, Dr. Richard Johnsson PhD, Pr<strong>of</strong>. Masad J Damha<br />
PhD, FCIC<br />
2:30 pm 47 Synthesis and evaluation <strong>of</strong> bicyclic ketal-based cationiclipids for the delivery<br />
<strong>of</strong> siRNA via lipid nanoparticle delivery systems<br />
Muthusamy Jayaraman, David Butler, Laxman Eltepu, Martin Maier, Tom<br />
Madden, Michael Hope, Ying Tam, Barbera Mui, Andrew Sprague, Akin<br />
Akinc, Soma De, G. Rajeev Kallanthottathil, Muthiah Manoharan<br />
2:45 pm 48 Development <strong>of</strong> a stability-indicating, ion-pair RP-HPLCmethod for separation<br />
and quantitative determination <strong>of</strong> two siRNA duplexes in aliposome<br />
Veeravagu Murugaiah, William Zedalis, Gary Lavine, Klaus Charrise, Muthiah<br />
Manoharan<br />
3:00 pm 49 Novel applications <strong>of</strong> aerosol-based detectors for theanalysis <strong>of</strong> nonchromophore,<br />
multi-lipid, drug delivery vehicles<br />
William Zedalis, Matthias Kretschmer, Muthiah Manoharan<br />
3:15 pm Intermission<br />
3:30 pm 50 Optimizing the LALassay for detection <strong>of</strong> bacterial endotoxin in conjugated<br />
and formulated siRNAs<br />
Mara Broberg, Kathy Mills, Klaus Charisse, William Zedalis, Muthiah<br />
Manoharan<br />
3:45 pm 51 Conjugationstrategies for RNAs using copper-catalyzed click chemistry<br />
Chang Geng Peng, Takeshi Yamada, Shigeo Matsuda, Haripriya Addepalli,<br />
Rowshon Alam, Narayanannair Jayaprakash, Muthusamy Jayaraman, David<br />
Butler, Rajendra Pandey, Kathy Mills, Martin Maier, Klaus Charrise, G.<br />
Rajeev Kallanthottathil, Muthiah Manoharan<br />
4:00 pm 52 Non-nucleoside building blocks for copper-assistedand copper-free click<br />
chemistry for synthesis <strong>of</strong> oligonucleotide conjugates<br />
K.N. Jayaprakash, Chang G. Peng, Takeshi Yamada, David Butler, G Rajeev<br />
Kallanthottathil, Martin Maier, Muthiah Manoharan
4:15 pm 53 Solid-support immobilized, reusable Cu (I)catalyst for "click reactions" <strong>of</strong><br />
oligonucleotides with ligands<br />
Laxman Eltepu, Chang G. Peng, K. Narayanannair Jayaprakash, Takeshi<br />
Yamada, Muthusamy Jayaraman, G. Rajeev Kallanthottathil, Muthiah<br />
Manoharan<br />
4:30 pm 54 Synthesis <strong>of</strong> oligo spermine-containingoligonucleotides for siRNA delivery<br />
Shigeo Matsuda, Gang Wang, Ligang Zhang, Tianlei Lei, Rowshon Alam,<br />
Chang G Pang, K. N. Jayaprakash, Takeshi Yamada, David Butler, Maria<br />
Frank-Kamenetsky, Martin Maier, Klaus Charrise, Kevin Fitzgerald, G. Rajeev<br />
Kallanthottathil, Muthiah Manoharan<br />
4:45 pm Concluding Remarks
CARB Todd Lowary Wednesday, August 25, 2010<br />
Oral<br />
Recognition <strong>of</strong> DNA: Recent Advances - AM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Cosponsored<br />
by:<br />
MEDI<br />
Organizers: Dev Arya<br />
Presiders: Dev Arya<br />
Duration: 8:00 am - 11:35 am<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
8:00 am Introductory Remarks<br />
8:05 am 55 Allosteric modulation <strong>of</strong> DNA by minor groove binding polyamides<br />
David M Chenoweth PhD, Pr<strong>of</strong>. Peter B Dervan PhD<br />
8:45 am 56 Strong and selective molecular recognition <strong>of</strong> the DNA minor groove:<br />
Compound and DNA chemistry and unusual conformational matching<br />
W. David Wilson, Rupesh Nanjunda, Arvind Kumar, Manoj Munde, Yang Liu,<br />
Rebecca Hunt, David W. Boykin<br />
9:25 am Intermission<br />
9:35 am 57 Toward DNA recognition by a Janus Wedge approach<br />
Pr<strong>of</strong>. Larry W McLaughlin PhD, Ayan Pal, Dr. Han Chen PhD, Dr. Meena<br />
Meena PhD
10:15 am 58 Recognition <strong>of</strong> DNA major groove<br />
Pr<strong>of</strong>. Dev P Arya, Dr. Liang Xue, Mr. Sunil Kumar<br />
10:55 am 59 Identification and cleavage site analysis <strong>of</strong> DNA sequences bound strongly by<br />
bleomycin<br />
Pr<strong>of</strong>. Sidney M Hecht PhD<br />
CARB Todd Lowary Wednesday, August 25, 2010<br />
Oral<br />
Recognition <strong>of</strong> DNA: Recent Advances - PM Session<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Dev Arya<br />
Presiders: Dev Arya<br />
Duration: 1:30 pm - 5:05 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
1:30 pm 75 Role <strong>of</strong> DNA topography in recognition by proteins and small molecules<br />
Pr<strong>of</strong>. Thomas D. Tullius<br />
2:10 pm 76 Selective modulation <strong>of</strong> DNA polymerase activity by fixed-conformation<br />
nucleoside analogs<br />
Pr<strong>of</strong>. Martin Egli, Dr. Robert L. E<strong>of</strong>f, Dr. Victor E. Marquez, Pr<strong>of</strong>. F. Peter<br />
Guengerich
2:50 pm 77 Structural mechanisms underlying DNA replication and human DNA mismatch<br />
repair<br />
Pr<strong>of</strong>. Lorena S. Beese PhD, Elizabeth McSweeney, Pr<strong>of</strong> Paul L Modrich PhD,<br />
Jillian Orans PhD, Quincy Tseng, Weina Wang<br />
3:30 pm Intermission<br />
3:45 pm 78 Manipulating the electrostatic potential in the DNA grooves: Effect on<br />
thermodynamic stability, ion binding, hydration and structure<br />
Pr<strong>of</strong>essor Barry Gold PhD<br />
4:25 pm 79 Sequence-dependent recognition <strong>of</strong> minor groove width<br />
Dr. Barry Honig
CARB Todd Lowary Wednesday, August 25, 2010<br />
Oral<br />
General Papers - AM Session<br />
Glycobiology<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 252A<br />
Organizers: Todd Lowary<br />
Presiders: Eugene Ci<strong>of</strong>fi<br />
Duration: 9:00 am - 11:35 am<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
9:00 am 60 Synthesis and characterization <strong>of</strong> chain-end functionalizable glycopolymer and its<br />
oriented glyco-macroligand formation<br />
Satya Nandana Narla, Xue-Long Sun PhD<br />
9:20 am 61 Separation and characterization <strong>of</strong> the glycosaminoglycan components <strong>of</strong><br />
proteoglycans<br />
Mellisa Ly, Tatiana N Laremore PhD, Kemal Solakyildirim MS, Pr<strong>of</strong>essor Robert<br />
J. Linhardt PhD<br />
9:40 am 62 Synthesis <strong>of</strong> nucleotide activated L-sugars from polyols via bioconversion<br />
Dr. Ryan D Woodyer PhD<br />
10:00 am Intermission<br />
10:15 am 63 Glycomorphology <strong>of</strong> the pulmonary vasculature: Endothelial cell glycocalyx and
endothelial barrier function<br />
Pr<strong>of</strong>essor Eugene Ci<strong>of</strong>fi<br />
10:35 am 64 Multidimensional glycan arrays for enhanced lectin and antibody pr<strong>of</strong>iling<br />
Dr. Yalong Zhang, Dr. Christopher Campbell MD, PhD, Dr. Qian Li, Dr. Jeffrey<br />
Gildersleeve<br />
10:55 am 65 Peptide nucleic acids bind strongly and sequence selectively to double helical<br />
RNA<br />
Mr Thomas T Zengeya, Dr Ming Li PhD, Dr Eriks Rozners PhD<br />
11:15 am 66 Post surface function method for preparation <strong>of</strong> liposomal glyco-conjugates<br />
MS Hailong Zhang, Dr. Yong Ma, Pr<strong>of</strong>. Xue-Long Sun<br />
CARB Todd Lowary Wednesday, August 25, 2010<br />
Oral<br />
General Papers - PM Session<br />
Glycobiology and Computation<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 252A<br />
Organizers: Todd Lowary<br />
Presiders: Steven Graham<br />
Duration: 1:00 pm - 3:55 pm
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
1:00 pm 67 Approach to study carbohydrate-carbohydrate interactions involved in myelin<br />
using glycolipids assay in microtiter plate<br />
Jingsha Zhao, Pr<strong>of</strong>. Amit Basu<br />
1:20 pm 68 WITHDRAWN<br />
1:40 pm 69 Synthesis and characterization <strong>of</strong> protein glycopolymer conjugate<br />
Valentinas Gruzdys, Xue-Long Sun PhD<br />
2:00 pm 70 Robust analytical development for oligonucleotide manufacture<br />
Dr. Ipsita Roymoulik<br />
2:20 pm Intermission<br />
2:35 pm 71 Glycomics studies on central nervous system<br />
Dr. Fuming Zhang, Dr. Zhenling Liu, Kemal Solakyildirim, Dennis Pu, Pr<strong>of</strong>.<br />
Robert J. Linhardt<br />
2:55 pm 72 Computational studies on the interactions <strong>of</strong> mannose with DOPC and POPC<br />
phospholipids<br />
Dr Parthasarathi R, Dr Gnanakaran S<br />
3:15 pm 73 Optimizatin <strong>of</strong> analysis <strong>of</strong> glycosaminoglycans in biological samples<br />
Boyangzi Li, Pr<strong>of</strong>. Robert J Linhardt PhD<br />
3:35 pm 74 Conformational analysis <strong>of</strong> nucleosides and nucleotides: PSEUROT 2010<br />
Steven M. Graham
CARB Todd Lowary Wednesday, August 25, 2010<br />
Poster<br />
General Posters - EVE Session<br />
Location: Boston Convention & Exposition Center<br />
Room: Hall C<br />
Organizers: Todd Lowary<br />
Duration: 8:00 pm - 10:00 pm<br />
Pub<br />
Presentation Title<br />
#<br />
80 Glycosylation <strong>of</strong> substituted 4H-1,2,4-triazole-3-thiol<br />
Pr<strong>of</strong>essor El Sayed H. El Ashry PhD,DSc<br />
81 Expression and characterization <strong>of</strong> enzymes for bioenzymatic synthesis <strong>of</strong> heparin<br />
Priscilla Paul, Wenjing Zhao, Pr<strong>of</strong>. Robert Linhardt, Pr<strong>of</strong>. Jonathan Dordick, Pr<strong>of</strong>. Jian Liu<br />
82 Catalytic conversion <strong>of</strong> biomass-derived carbohydrates to useful chemicals in one step<br />
Dr. Weiran Yang, Pr<strong>of</strong>. Ayusman Sen<br />
83 Behaviour <strong>of</strong> polysaccharide aggregates in asymmetricalfield-flow fractionation and sizeexclusion<br />
chromatography<br />
Leena Pitkänen, Maija Tenkanen, Päivi Tuomainen<br />
84 Key new observations in the synthesis <strong>of</strong> thiosialosides<br />
Ms Ines F Oliveira, Dr Goreti R Morais PhD, Mr Bradley R Springett, Dr Robert A<br />
Falconer PhD<br />
85 Synthesis and structural optimization <strong>of</strong> antifungal kanamycin B analogs<br />
Marina Y FOSSO, Yukie KAWASAKI, Sanjib SHRESTHA, Jon TAKEMOTO, Tom C.-<br />
W. CHANG<br />
86 E. coli K5 heparosan production for bioengineered heparin
Zhenyu Wang, Mellisa Ly, Fuming Zhang, Zhenqing Zhang, Jonathan S. Dordick, Robert J.<br />
Linhardt<br />
87 Stochastic simulation <strong>of</strong> lectin microarrays with nanosensor transducers: Potential platforms<br />
for optimal, high-throughput screening and pr<strong>of</strong>iling <strong>of</strong> glycoproteins<br />
Mr. Nigel F Reuel, Dr. Jin-Ho Ahn PhD, Dr. Michael S. Strano PhD<br />
88 Evaluation <strong>of</strong> different thioesters for glycocluster synthesis applying native chemical<br />
ligation<br />
Johannes W. Wehner, Pr<strong>of</strong>. Dr. Thisbe K. Lindhorst<br />
89 NMR for structure elucidation <strong>of</strong> commercially available heparin polysaccharides<br />
Kemal Solakyildirim, Scott A. McCallum, Robert J. Linhardt<br />
90 Syntheses <strong>of</strong> C-5-spirocyclic C-glycoside SGLT2 inhibitors<br />
Benjamin A. Thuma, Dr. Vincent Mascitti PhD, Dr. Ralph P. Robinson PhD, Cathy<br />
Préville, Matthew R. Reese, Dr. Robert J. Maguire PhD, Christopher L. Carr, Michael T.<br />
Leininger, André Lowe, Claire M. Steppan<br />
91 Co-axial cellulose nan<strong>of</strong>ibers for electrical applications<br />
Minoru Miyauchi, Dr. Jianjun Miao PhD, Dr. Trevor J. Simmons PhD, Pr<strong>of</strong>. Jonathan S.<br />
Dordick PhD, Pr<strong>of</strong>. Robert J. Linhardt PhD<br />
92 Study on therelative reactivity <strong>of</strong> glycosyl acceptors in the glycosylations <strong>of</strong>2-Azido-2deoxy-galactosides<br />
Jane Kalikanda, Dr. Zhitao Li<br />
93 Synthesis <strong>of</strong> tailored glycoconjugates for the precise detection <strong>of</strong> pathogens<br />
Ashish A. Kulkarni, Dr. Suri S. Iyer<br />
94 Structure-activity studies <strong>of</strong> synthetic glycophosphatidylinositol anchored proteins<br />
Dr. Carl V Christianson, Pr<strong>of</strong>. Peter H Seeberger<br />
95 Chemoenzymatic synthesis <strong>of</strong> heparan sulfate<br />
Dr Renpeng Liu, Pr<strong>of</strong> Jian Liu PhD, Dr Yongmei Xu<br />
96 Synthesis <strong>of</strong> a fluorous-tagged disaccharide for the enzymatic preparation <strong>of</strong> heparin<br />
oligosaccharides
Sayaka Masuko, Smritilekha Bera, Robert J. Linhardt<br />
97 Preventing the transmission <strong>of</strong> Plasmodium falciparum through the inhibition <strong>of</strong> malaria<br />
protein binding to placental chondroitin sulfate A<br />
Julie M Beaudet, Leandra J Mansur, Bo Yang, Fuming Zhang, Robert J Linhardt<br />
98 Real-time assessment <strong>of</strong> the morphological change <strong>of</strong> cellulose in response to enzymatic<br />
treatment<br />
Chi Nguyen<br />
99 Chemoselective glycosylation <strong>of</strong> hemoglobin as a potential oxygen therapeutic<br />
Thomas J Styslinger, Ning Zhang, Andre F. Palmer PhD, Peng G. Wang PhD<br />
100 Immobilization <strong>of</strong> enzymes relevant to bioengineered heparin synthesis<br />
Eric R. Sterner, Dr. Robert J. Linhardt, Dr. Jonathan S. Dordick, Dr. Jian Liu, Dr. Fuming<br />
Zhang, Wenjing Zhao, Priscilla Paul, Jeff Martin<br />
101 Analyses <strong>of</strong> anti Tn-antigen MLS128 monoclonal antibody binding to two or three<br />
consecutive Tn-antigen clusters by surface plasmon resonance (SPR) and NMR<br />
Ayano Takasaki-Matsumoto Ph.D., Shinya Hanashima Ph.D., Ami Aoki, Yoshiki<br />
Yamaguchi Ph.D., Reiko Sato, Hiroko Kawakami Ph.D., Mamoru Mizuno Ph.D., Pr<strong>of</strong>.<br />
Yoko Fujita-Yamaguchi Ph.D.<br />
102 Modular glycoconjugate tool set for assembly and presentation <strong>of</strong> multivalent carbohydrate<br />
ligands on surfaces<br />
Irene Abia, Brian Sanders, Michael D. Best, David C. Baker<br />
103 Analysis<strong>of</strong> the absorption <strong>of</strong> low molecular weight heparin in human umbilical cord tissue<br />
as a model for the prevention <strong>of</strong> cancer metastasis<br />
Amanda M. Weyers, Thangriala Sudha, Bo Yang, Boyangzi Li, Majde Takieddin, Fuming<br />
Zhang, Shaker A. Mousa, Robert J. Linhardt<br />
104 Synthesis and biological evaluation <strong>of</strong> a Gal(α1-2)GalCer analog<br />
Yanke Liang, Pr<strong>of</strong>. Amy R Howell PhD<br />
105 Thiol-click chemistry approach to glycomimetics: Novel stereoselective synthesis <strong>of</strong> (1-3)-<br />
S-thiodisaccharides<br />
Pr<strong>of</strong>. Zbigniew J. Witczak, Irena Bak-Sypien
106 High-throughput glycoarray for monitoring immune responses to a cancer vaccine<br />
Dr. Christopher Campbell M.D., Ph.D., Dr. Yalong Zhang Ph.D., Dr. Olaf Ludek Ph.D.,<br />
Mr. David Farnsworth, Dr. Jeffrey Gildersleeve Ph.D.<br />
107 NMR spectroscopic studies <strong>of</strong> APF: A small glycopeptide possessing potent<br />
antiproliferative activity<br />
Dr Kristie M Adams PhD, Dr Piotr Kaczmarek PhD, Dr Susan K Keay MD, PhD, Dr<br />
Joseph J Barchi, Jr PhD<br />
108 Structural and quantitative analysis <strong>of</strong> disaccharides using CE with LIF detection<br />
Yuqing Chang, Tatiana Laremore, Fuming Zhang, Robert J Linhardt<br />
109 Development <strong>of</strong> a novel cancer vaccine based on multivalent presentation <strong>of</strong> tumorassociated<br />
carbohydrate antigens on gold nanoparticle scaffolds<br />
Raymond P Brinas Ph.D., Andreas Sundgren Ph.D., Micah Maetani, Omar Abbudayyeh,<br />
Howard A Young Ph.D., Michael Sanford, Joseph J Barchi Ph.D.<br />
110 Carbon nanotubes and chitosan as possible scaffolds for bone tissue regeneration<br />
Julia Stone, Yetunde Olusanya, Whitney Jones, Pasakorn Traisawatwong, Melisa Stewart,<br />
Cordella Kelly-Brown, Laura Carson Ph.D., Aderemi Oki Ph.D., E. Gloria C. Regisford<br />
Ph.D.<br />
111 Biosynthesis <strong>of</strong> heparin by metabolic engineering <strong>of</strong> Chinese hamster ovary cells<br />
Leyla Gasimli, Jongyoun Baik PhD, Dr. Susan Sharfstein PhD, Pr<strong>of</strong>. Robert J. Linhardt<br />
PhD<br />
112 Purification strategies for separation <strong>of</strong> capsular polysaccharide from fermentation broth<br />
Ujjwal Bhaskar, Zhenyu Wang, Dr. Fuming Zhang, Pr<strong>of</strong>. Jonathan S. Dordick PhD, Pr<strong>of</strong>.<br />
Robert J. Linhardt PhD<br />
113 Design, synthesis, and characterization <strong>of</strong> sulfated N-aryl aminoglycosides<br />
Amanda M. Fenner, Robert J. Kerns PhD<br />
114 Photo-click immobilization <strong>of</strong> carbohydrates on polymeric surfaces: An effortless method to<br />
functionalize surfaces for biomolecular recognition studies<br />
Oscar Norberg, Lingquan Deng, Mingdi Yan, Ol<strong>of</strong> Ramström
115 On-line micr<strong>of</strong>low high-performance liquid chromatography with nano-electrospray<br />
ionization mass spectrometry for heparan sulfate disaccharide analysis<br />
Dr. Bo Yang, Kemal Solakyildirim, Jeffrey G. Martin, Tatiana Laremore, Pr<strong>of</strong>. Robert J.<br />
Linhardt
CARB Todd Lowary Thursday, August 26, 2010<br />
Oral<br />
General Papers - AM Session<br />
Synthetic <strong>Chemistry</strong><br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Todd Lowary<br />
Presiders: Cecilia Marzabadi<br />
Duration: 8:45 am - 12:00 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
8:45 am 116 De novo synthesis <strong>of</strong> a 2-acetamido-4-amino-2,4,6-trideoxy-D-galactose (AAT)<br />
building block for the preparation <strong>of</strong> the zwitterionic polysaccharide A1 (PS A1)<br />
repeating subunit <strong>of</strong> Bacteroides fragilis<br />
Dr. Rajan Pragani PhD, Pr<strong>of</strong>. Dr. Peter H Seeberger PhD<br />
9:05 am 117 Automated solution-phase synthesis <strong>of</strong> alpha-galactosides<br />
Rajarshi Roychoudhury, Pr<strong>of</strong>. Nicola L. B. Pohl<br />
9:25 am 118 Study <strong>of</strong> the protecting group participation and stereoselectivity <strong>of</strong> 2-azido-2deoxygalactopyranosyl<br />
donors<br />
Pr<strong>of</strong>. Zhitao Li<br />
9:45 am 119 Reactions <strong>of</strong> glycals with ortho-substituted benzanilines<br />
Pr<strong>of</strong>. Cecilia H Marzabadi, Katherine Kochalski<br />
10:05 am 120 Novel glycolipids in CD1d-mediated immunity: Synthesis <strong>of</strong> new agonists for<br />
CD1d
Justyna Wojno, John-Paul Jukes, Paolo Polzella, Vincenzo Cerundolo, Liam R<br />
Cox, Gurdyal S Besra<br />
10:25 am Intermission<br />
10:40 am 121 Automated synthesis <strong>of</strong> systematic di- and tri-saccharide libraries<br />
Xin Liu, Dr. Beatrice Y. M. Collet, Shu-Lun Tang, Sahana K. Nagappayya,<br />
Rajarshi Roychoudhury, Dr. Xueshu Li, Heather Edwards, Pr<strong>of</strong>. Nicola L. B. Pohl<br />
11:00 am 122 Probe <strong>of</strong> activated glycoside building block stability for automated solution-phase<br />
synthesis <strong>of</strong> carbohydrate libraries<br />
Dr. Beatrice Y. M. Collet, Xin Liu, Shu-Lun Tang, Heather Edwards, Sahana<br />
Nagappayya, Lin Liu, Pr<strong>of</strong>. Nicola L. B. Pohl<br />
11:20 am 123 Synthesis <strong>of</strong> unnatural glycosaminoglycans and evaluation <strong>of</strong> their interaction<br />
with proteins<br />
Dr. Smritilekha Bera PhD<br />
11:40 am 124 Grafting <strong>of</strong> cellulose esters by single electron transfer living radical<br />
polymerization Set-LrP<br />
Dr Petr Vlcek, Vladimir Raus, Dr Miroslav Janata, Dr Jaroslav Kriz, Petra<br />
Latalova, Eva Cadova<br />
CARB Todd Lowary Thursday, August 26, 2010<br />
Oral<br />
General Papers - PM Session<br />
Polysaccharides<br />
Location: Boston Convention & Exhibition Center<br />
Room: Room 251<br />
Organizers: Todd Lowary<br />
Presiders: Ravin Narain
Duration: 1:00 pm - 4:55 pm<br />
Pres<br />
Time<br />
Pub<br />
#<br />
Presentation Title<br />
1:00 pm 125 Solvation and hydrolysis <strong>of</strong> cellulose with transition metal salts<br />
Dr. Veronika Viner, Dr. Benjamin G. Harvey, Dr. Roxanne L Quintana<br />
1:20 pm 126 <strong>Chemical</strong> derivatization <strong>of</strong> glucan microparticles for targeted delivery<br />
Dr. Ernesto R Soto, Dr. Gary R Ostr<strong>of</strong>f<br />
1:40 pm 127 Degradation mechanism and crystal structure change <strong>of</strong> cellulose during TEMPO-<br />
NaOCl-NaBr selective oxidation and its biomedical application as hemostatics<br />
Dr.,Associate Pr<strong>of</strong>. Bin Sun PhD, Wenxiu Han, Wei Wang, Chunju Gu, Jinhong<br />
Ma, Meifang Zhu, Borun Liang<br />
2:00 pm 128 Temperature-dependent chain-collapse behavior <strong>of</strong> cellulose ethers in dilute<br />
solution<br />
Hongwei Shen, Robert Sammler, David Redwine, David Meunier, Meinolf<br />
Brackhagen<br />
2:20 pm 129 Improving dyeability <strong>of</strong> nylon wiith nano-chitosan<br />
Ms Ho Yan Wong, Ms Hoi Ying Lui, Ms Li Wei Liu, Dr Yau Shan Szeto PhD<br />
2:40 pm 130 Modeling <strong>of</strong> Congo red adsorption on a surface <strong>of</strong> crystalline cellulose using<br />
molecular dynamics<br />
Dr. Miroslaw Wyszomirski, Dr. Karim Mazeau<br />
3:00 pm Intermission<br />
3:15 pm 131 Evaluation <strong>of</strong> glycopolymers and glyconanoparticles for biosensing and gene<br />
delivery applications<br />
Pr<strong>of</strong>. Ravin Narain PhD, Miss Marya Ahmed<br />
3:35 pm 132 Application <strong>of</strong> data mining techniques to differentiate glucose-containing<br />
disaccharide ions fragmented via infrared-multiple photon dissociation using<br />
tunable lasers and Fourier transform ion cyclotron resonance mass spectrometry<br />
Dr. Sarah Stefan PhD, Mohammad U. Ehsan, Dr. Alexander Aksenov PhD, Dr.
3:55 pm 133<br />
Vladimir Boginksi PhD, Dr. Brad Bendiak PhD, Dr. John Eyler PhD<br />
Influence <strong>of</strong> the structure <strong>of</strong> phosphoramidates on flame retardant properties <strong>of</strong><br />
cellulose<br />
Victoria Salimova, Dr. Sabyasachi Gaan, Dr. Joëlle Grützmacher<br />
4:15 pm 134 High-flux thin-film nan<strong>of</strong>ibrous composite ultrafiltration membranes containing<br />
cellulose barrier layer<br />
Dr. Hongyang Ma, Pr<strong>of</strong>. Benjamin S. Hsiao, Pr<strong>of</strong>. Benjamin Chu<br />
4:35 pm 135 Discovery <strong>of</strong> 6-deoxy-D-altrose in nature<br />
Pr<strong>of</strong>essor Masakuni Tako PhD, Takuya Yogi, Pr<strong>of</strong>essor Ken Izumori PhD,<br />
Pr<strong>of</strong>essor Hideharu Ishida PhD, Pr<strong>of</strong>essor Makoto Kiso PhD
CARB 1<br />
Developing an enzymatic approach to synthesize heparan sulfates<br />
Jian Liu (1) , jian_liu@unc.edu, Eshelman School <strong>of</strong> Pharmacy, Chapel Hill North<br />
Carolina 27599, United States . (1) Med Chem and Nat Prod, University <strong>of</strong> North<br />
Carolina, Chapel Hill North Carolina 27599, United States<br />
Heparan sulfate is a sulfated glycan that exhibits essential physiological<br />
functions. Interrogation <strong>of</strong> the specificity <strong>of</strong> heparan sulfate-mediated activities<br />
demands a library <strong>of</strong> structurally defined oligosaccharides. Synthesis <strong>of</strong> heparan<br />
sulfate using enzymes provides a promising approach because <strong>of</strong> the high<br />
regioselectivity <strong>of</strong> heparan sulfate biosynthetic enzymes. The synthesis <strong>of</strong><br />
heparan sulfate involves up to 15 specialized enzymes, including<br />
sulfotransferases, an epimerase and glycosyltransferases. All <strong>of</strong> these enzymes<br />
have been expressed effectively in bacteria and in yeast in our lab. Using these<br />
enzymes, we are able to synthesize oligosaccharides with different sulfation<br />
patterns and sizes. Furthermore, we demonstrated the feasibility <strong>of</strong> engineering<br />
the substrate specificity <strong>of</strong> heparan sulfate sulfotransferases using a structurally<br />
guided mutagenesis approach. The engineered enzymes have provided us the<br />
opportunity to prepare the sulfated polysaccharides that can not be achieved by<br />
the wild type enzymes. Overall, our method <strong>of</strong>fers a generic for developing<br />
carbohydrate-based therapeutics.<br />
CARB 2<br />
Creating and implementing breakthrough medical technology<br />
Robert S Langer (1) , rlanger@mit.edu, 45 Carleton Street, Cambridge MA 02142,<br />
United States . (1) Massachusetts Institute <strong>of</strong> Technology, United States<br />
Several different case studies in the areas <strong>of</strong> drug delivery, medical devices and<br />
biotherapeutics will be discussed. Each study will be examined in terms <strong>of</strong> the<br />
process and excitement <strong>of</strong> discovery, initial resistance by the scientific<br />
community to the discovery in some cases, the way very broad patents were<br />
received, how the technologies were transferred to companies, and the way they<br />
have been or are trying to be commercialized. Among those that will be<br />
discussed are studies involving heparinase and heparin in which Bob Linhardt<br />
played a seminal role.<br />
CARB 3<br />
Advances in heparin analysis and synthesis<br />
Robert J. Linhardt (1) , linhar@rpi.edu, Center for Biotechnology and<br />
Interdisciplinary Studies, 110 8th Street, Troy NY 12180, United States . (1)
Departements <strong>of</strong> <strong>Chemistry</strong>, Biology and <strong>Chemical</strong> Engineering, Rensselaer<br />
Polytechnic Institute, Troy NY 12180, United States<br />
Heparin, a member <strong>of</strong> the heparan sulfate (HS) family <strong>of</strong> proteoglycans (PGs), is<br />
an important therapeutic agent.<br />
Disaccharide analysis on HS requires 10 4 cells using LC-MS and oligosaccharide<br />
mapping affords critical structural information. Sequencing <strong>of</strong><br />
oligo/polysaccharides is now possible using FTICR-MS with EDD leading to<br />
structure-activity relationships (SAR). Improved SAR facilitates new drug<br />
development, safer heparins, and has led to an early understanding <strong>of</strong> the PGome<br />
and its importance in ESC differentiation. Novel approaches are currently<br />
underway in our laboratory to explore HS biosynthesis, including the metabolic<br />
engineering <strong>of</strong> CHO cells and the construction <strong>of</strong> an artificial Golgi on a digital<br />
micr<strong>of</strong>luidic platform. Finally, research is now being directed to develop largescale<br />
enzyme-assisted synthesis <strong>of</strong> HS/heparin aimed at replacing animal<br />
sourced pharmaceutical heparin with a bioengineered product.<br />
CARB 4<br />
Development <strong>of</strong> a fully synthetic three-component carbohydrate-based<br />
cancer vaccine<br />
Geert-Jan Boons (1) , gjboons@ccrc.uga.edu, 315 Riverbend Rd, Athens GA<br />
30606, United States . (1) complex <strong>Carbohydrate</strong> Research Center, University <strong>of</strong><br />
Georgia, Athens Georgia 30606, United States<br />
The mucin MUC1 is over-expressed by as much as 50-fold and aberrantly<br />
glycosylated by the<br />
majority <strong>of</strong> carcinomas. Recently, a National Cancer Institute translational<br />
research working group determined that MUC1 is one <strong>of</strong> the most promising<br />
targets for cancer vaccine development. However, a MUC1-based cancer<br />
vaccine that can elicit robust humoral and cellular immune responses has not<br />
been reported. We show here that an appropriately glycosylated MUC1 peptide<br />
covalently linked to promiscuous helper T-epitope and a Toll-like receptor (TLR)<br />
agonist can elicit robust humoral and cellular immune responses and was<br />
efficacious in reversing tolerance and generating a therapeutic response in a<br />
mouse model <strong>of</strong> mammary cancer. The superior properties <strong>of</strong> the vaccine<br />
candidate are attributed to the local production <strong>of</strong> cytokines, upregulation <strong>of</strong> costimulatory<br />
proteins, enhanced uptake by macrophages and dendritic cells, and<br />
avoidance <strong>of</strong> epitope suppression.<br />
CARB 5<br />
Chemoenzymatic synthesis and antibody recognition <strong>of</strong> HIV-1 V3<br />
glycopeptides
Lai-Xi Wang (1) , lwang@som.umaryland.edu, 725 W. Lombard Street, Baltimore<br />
MD 21201, United States . (1) Institute <strong>of</strong> Human Virology and Department <strong>of</strong><br />
Biochemistry & Molecular Biology, University <strong>of</strong> Maryland School <strong>of</strong> Medicine,<br />
Baltimore MD 21201, United States<br />
A typical HIV-1 gp120 envelope glycoprotein carries more than 20 N-glycans.<br />
These oligosaccharides are essential for the virus to evade immune response<br />
and meanwhile can serve as ligands for dendritic cell-mediated HIV-1<br />
transmission. The third variable domain (V3) <strong>of</strong> gp120 is a predominant<br />
neutralization determinant that carries three conserved N-glycans within or<br />
around the loop. To elucidate the roles <strong>of</strong> these N-glycans in antibody<br />
recognition, we have synthesized a series <strong>of</strong> homogeneous, full-size V3<br />
glycopeptides by a chemoenzymatic approach and evaluated their binding to two<br />
broadly neutralizing antibodies: the V3-speicific antibody 447-52D and the<br />
carbohydrate-specific antibody 2G12. Our studies indicate that the complex type<br />
N-glycan at N301 has moderate effects on the recognition <strong>of</strong> V3 to 447-52D,<br />
while the V3 domain facilitates the binding <strong>of</strong> antibody 2G12 to the two highmannose<br />
type N-glycans at the V3 base. This study provides new insights in<br />
HIV-1 vaccine design.<br />
CARB 6<br />
Entirely carbohydrate vaccine constructs and their application in probing<br />
glycoimmunology<br />
Peter R Andreana (1) , pra@chem.wayne.edu, 5101 Cass Ave, Detroit MI 48202,<br />
United States . (1) <strong>Chemistry</strong>, Wayne State University, Detroit MI 48202, United<br />
States<br />
For effective immunity, a strong and long-term response MUST be generated<br />
through both cellular and humoral arms <strong>of</strong> the immune system, involving class II<br />
major histocompatibility complex (MHCII) proteins, CD4 + T-cells and B-cells in a<br />
T-cell-dependent cascade. A new class <strong>of</strong> bacterial polysaccharides,<br />
characterized by an alternating zwitterionic charge motif on adjacent<br />
monosaccharides, has been shown to stimulate T- and B-cell immune responses<br />
effectively. We hypothesize that chemically conjugating tumor-associated<br />
carbohydrate antigens (TACAs) to zwitterionic polysaccharides (ZPSs) will lead<br />
to carbohydrate vaccines that can stimulate strong immunity.<br />
A process for evaluating immunogenicity, specificity, and function <strong>of</strong> the novel<br />
construct will include isolation, purification, chemical modification <strong>of</strong> PS A1 and<br />
subsequent in vivo mouse studies. ELISA, FACS and cytotoxicity studies will<br />
reveal that an immune response is specific for the conjugated Tn antigen and<br />
that cell killing is achieved using a complement dependent approach.
CARB 7<br />
<strong>Carbohydrate</strong> polymer assembly: How do mycobacteria do it?<br />
Laura L. Kiessling (1) , kiessling@chem.wisc.edu, 1101 University Ave., Madison<br />
WI 53706, United States . (1) Departments <strong>of</strong> <strong>Chemistry</strong> and Biochemistry,<br />
University <strong>of</strong> Wisconsin-Madison, Madison WI 53706, United States<br />
<strong>Carbohydrate</strong> polymers are the most abundant organic compounds on this<br />
planet. They mediate fundamental processes in humans but also can be<br />
essential for pathogen survival. A key component <strong>of</strong> the mycobacterial cell wall is<br />
a polysaccharide that contains galact<strong>of</strong>uranose (Galf) residues. Because Galf<br />
residues are not constituents <strong>of</strong> human glycoproteins or glycolipids, the enzymes<br />
mediating their incorporation should serve as antimicrobial targets. Blocking<br />
Mycobacterium tuberculosis, the causative agent <strong>of</strong> tuberculosis (TB), would be<br />
valuable because TB results in approximately 2 million deaths/year. To this end,<br />
we are exploring the synthesis <strong>of</strong> the galactan portion <strong>of</strong> the cell wall. One key<br />
enzyme in this pathway is the bifunctional glycosyltransferase, GlfT2, that<br />
assembles a polysaccharide composed <strong>of</strong> alternating 1,5- and 1,6-linked Galf<br />
residues. Our investigations were driven by several questions: How does a single<br />
glycosyltransferase generate both types <strong>of</strong> Galf linkages? Does the enzyme have<br />
multiple active sites? Does elongation occur by a process that is distributive or<br />
processive? How is polysaccharide length controlled in the absence <strong>of</strong> a<br />
template? Can inhibitors that block this enzyme be found? We have explored<br />
these questions using a range <strong>of</strong> approaches that span biology and chemistry,<br />
and our results will be discussed. We anticipate that the strategies developed to<br />
investigate the molecular mechanism <strong>of</strong> GlfT2 can be readily applied to probe a<br />
wide variety <strong>of</strong> enzymatic polymerization reactions, including those that result in<br />
polysaccharide production.<br />
CARB 8<br />
Lipopolysaccharide transport and assembly in Escherichia coli
Daniel Kahne (1) , kahne@chemistry.harvard.edu, 12 Oxford St, Cambridge MA<br />
02138, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology,<br />
Harvard University, Cambridge MA 02138, United States<br />
The outer membrane <strong>of</strong> Gram-negative bacteria contains an outer leaflet<br />
composed <strong>of</strong> lipopolysaccharide (LPS) that is transported to this location by a<br />
pathway that is essential for viability. It has been suggested that inhibitors <strong>of</strong> this<br />
pathway could be useful antibiotics. In Escherichia coli, eight essential proteins<br />
have been identified to function in the proper assembly <strong>of</strong> LPS following its<br />
biosynthesis. This assembly process involves release <strong>of</strong> LPS from the inner<br />
membrane, transport across the periplasm, and insertion into the outer leaflet <strong>of</strong><br />
the outer membrane. I will talk about the mechanism <strong>of</strong> LPS transport and<br />
assembly and the development <strong>of</strong> tools that could lead to the discovery <strong>of</strong><br />
inhibitors <strong>of</strong> this process.<br />
CARB 9<br />
Arrays and automated oligosaccharide synthesis<br />
Nicola L. B. Pohl (1) , npohl@iastate.edu, 2756 Gilman, Ames IA 50011, United<br />
States . (1) Department <strong>of</strong> <strong>Chemistry</strong> and The Plant Sciences Institute, Iowa<br />
State University, Ames IA 50011, United States<br />
Practical methods to enable the automated solid-phase synthesis <strong>of</strong> peptides and<br />
nucleic acids have been crucial to driving genomics and proteomics.<br />
Unfortunately, solid-phase methods usually require large excesses <strong>of</strong> building<br />
blocks to achieve the high yields needed when intermediates cannot be purified<br />
and therefore are essentially limited to building blocks that are made in a few<br />
steps. This talk will discuss the development <strong>of</strong> an alternative solution-phase<br />
automation strategy to iterative synthesis that is based on soluble fluorocarbon<br />
tags and the interface <strong>of</strong> this fluorous synthesis strategy with the formation <strong>of</strong><br />
small molecule microarrays. The scope and current limitations <strong>of</strong> this method will<br />
also be presented.<br />
CARB 10<br />
Bioactive N-linked glycopeptides and glycoprotein conjugates<br />
Thomas J. Tolbert (1) , tolbert@indiana.edu, 800 E. Kirkwood Ave., <strong>Chemistry</strong><br />
Building, Bloomington IN 47405, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong>,<br />
Indiana University, Bloomington IN 47405, United States<br />
N-linked glycosylation is a highly prevalent posttranslational modification that is<br />
also present in a large fraction <strong>of</strong> FDA approved protein therapeutics. We have<br />
recently developed methods for the synthesis <strong>of</strong> homogeneous N-linked<br />
glycopeptides (1) and glycoproteins (2) and are applying these techniques to the
study <strong>of</strong> bioactive glycopeptides and semisynthetic glycoprotein conjugates.<br />
These studies will be presented with a focus on the effects <strong>of</strong> N-linked<br />
glycosylation on diabetes related peptides and glycosylated antibody fragments.<br />
N-linked glycosylation has proven to have some unexpected effects on the<br />
physical properties and bioactivity <strong>of</strong> glycoconjugates in these systems that may<br />
be useful in the development <strong>of</strong> glycopeptide and glycoprotein therapeutics.<br />
(1) Journal <strong>of</strong> the <strong>American</strong> <strong>Chemical</strong> <strong>Society</strong> 2010, 132, 3211-3216.<br />
(2) Journal <strong>of</strong> the <strong>American</strong> <strong>Chemical</strong> <strong>Society</strong> 2009, 131, 13616-13618.<br />
CARB 11<br />
Efficient chemoenzymatic synthesis <strong>of</strong> sialyl Lewis antigens using<br />
sialyltransferase mutants<br />
Xi Chen (1) , chen@chem.ucdavis.edu, One Shields Avenue, Davis CA 95616,<br />
United States ; Go Sugiarto (1) ; Kam Lau (1) ; Lei Zhang (1) ; Shengshu Huang (1) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong>, University <strong>of</strong> Califorina-Davis, Davis California 95616,<br />
United States<br />
Sialic acids are a family <strong>of</strong> nine-carbon monosaccharides mainly found as the<br />
terminal residues <strong>of</strong> glycans on cell surfaces (1). More than 50 structurally<br />
distinct sialic acids have been found in nature (2, 3). Sialyl Lewis x is an important<br />
sialic acid-containing carbohydrate epitope involved in many biological processes<br />
such as inflammation (4) and cancer metastasis (5). Sialyl Lewis a is also involved<br />
in cancer progression (6). One <strong>of</strong> our research goals is to develop efficient<br />
chemoenzymatic method for the synthesis <strong>of</strong> sialyl Lewis x and sialyl Lewis a<br />
oligosaccharides containing structurally diverse sialic acid residues.<br />
The general biosynthetic pathway <strong>of</strong> sLe x and sLe a involves sialylation followed<br />
by fucosylation. In order to obtain sLe x and sLe a containing structurally diverse<br />
sialic acids, a more efficient approach would be to carry out the fucosylation<br />
before the sialylation so that different sialic acid forms can be introduced in the<br />
last glycosylation step. However, most sialyltransferases reported can not<br />
tolerate fucosylated substrates except for a viral alpha2,3-sialyltransferase (v-<br />
ST3Gal I) from myxoma virus (7). We have successfully cloned and expressed<br />
the v-ST3Gal I in E. coli. The enzyme has been used in the sialylation <strong>of</strong> Lewis x<br />
antigen. In addition, mutants <strong>of</strong> Pasteurella multocida sialyltransferase 1<br />
(PmST1), a multifunctional alpha2,3-sialyltransferase that has excellent<br />
expression level, solubility, and activity towards non-fucosylated substrates, have<br />
been designed and generated for direct sialylation <strong>of</strong> fucosylated<br />
oligosaccharides.<br />
1. Varki, A. (1997) Sialic acids as ligands in recognition phenomena, FASEB J.<br />
11, 248–255.<br />
2. Angata, T., and Varki, A. (2002) <strong>Chemical</strong> diversity in the sialic acids and
elated alpha-keto acids: an evolutionary perspective, Chem. Rev. 102, 439–469.<br />
3. Schauer, R. (2000) Achievements and challenges <strong>of</strong> sialic acid research,<br />
Glycoconjugate J. 17, 485–499.<br />
4. Rosen, S. D. (2004) Ligands for L-selectin: homing, inflammation, and beyond,<br />
Annu. Rev. Immunol. 22, 129–156.<br />
5. Fukuda, M. (1996) Possible roles <strong>of</strong> tumor-associated carbohydrate antigens,<br />
Cancer Res. 56, 2237-2244.<br />
6. Ugorski, M., and Laskowska, A. (2002) Sialyl Lewis(a): a tumor-associated<br />
carbohydrate antigen involved in adhesion and metastatic potential <strong>of</strong> cancer<br />
cells, Acta Biochim. Pol. 49, 303–311.<br />
7. Sujino, K., Jackson, R. J., Chan, N. W., Tsuji, S., and Palcic, M. M. (2000) A<br />
novel viral alpha2,3-sialyltransferase (v-ST3Gal I): transfer <strong>of</strong> sialic acid to<br />
fucosylated acceptors, Glycobiology 10, 313–320.<br />
CARB 12<br />
Synthesis <strong>of</strong> saponin vaccine adjuvants for immunotherapy<br />
David Y Gin (1) , gind@mskcc.org, 1275 York Ave., Box 379, New York NY 10065,<br />
United States . (1) Department <strong>of</strong> Molecular Pharmacology and <strong>Chemistry</strong>,<br />
Memorial Sloan-Kettering, New York NY 10065, United States<br />
Saponin natural products constitute some <strong>of</strong> the most promising adjuvants for<br />
immune response potentiation and dose-sparing in several experimental vaccine<br />
therapies. For example, melanoma, breast cancer, small cell lung cancer,<br />
prostate cancer, HIV-1, and malaria are among the numerous maladies targeted<br />
in vaccine clinical trials using Quillaja saponins as critical adjuvants for immune<br />
response augmentation. Recent synthetic efforts directed at the chemical<br />
synthesis and evaluation <strong>of</strong> adjuvant-active natural and non-natural saponins will<br />
be described.<br />
CARB 13<br />
Glycosylation engineering <strong>of</strong> human IgG-Fc for functional studies<br />
Lai-Xi Wang (1) , lwang@som.umaryland.edu, 725 W. Lombard St, Baltimore MD<br />
21201, United States . (1) Institute <strong>of</strong> Human Virology and Department <strong>of</strong><br />
Biochemistry & Molecular Biology, University <strong>of</strong> Maryland School <strong>of</strong> Medicine,<br />
Baltimore MD 21201, United States<br />
Monoclonal antibodies (MAbs) <strong>of</strong> the immunoglobulin G (IgG) type are an<br />
important class <strong>of</strong> therapeutic glycoproteins. Compelling evidence has indicated<br />
that the fine structures <strong>of</strong> the conserved N-glycan attached to the Fc domain are<br />
responsible for the effector functions <strong>of</strong> MAbs, including antibody-dependent<br />
cellular cytotoxicity (ADCC), complement-dependent cytotoxicity (CDC), and<br />
activation <strong>of</strong> apoptosis. However, the structural heterogeneity <strong>of</strong> Fc glycosylation
poses a big challenge in understanding the functional roles <strong>of</strong> IgG-Fc<br />
glycosylation. In this paper, we describe a chemoenzymatic method for IgG-Fc<br />
glycosylation engineering that involves expression <strong>of</strong> human IgG-Fc in yeast and<br />
subsequent glycan remodelling through endoglycosidase-catalyzed<br />
transglycosylation. This method permits the synthesis <strong>of</strong> various homogeneous<br />
glyc<strong>of</strong>orms <strong>of</strong> human IgG-Fc. The affinity <strong>of</strong> different glyc<strong>of</strong>orms to key Fc<br />
receptors is being evaluated by SPR analysis and will be discussed.<br />
CARB 14<br />
Chemoenzymatic synthesis <strong>of</strong> GPI-anchored proteins and glycoproteins<br />
Zhongwu Guo (1) , zwguo@chem.wayne.edu, 5101 Cass Avenue, Detroit<br />
Michigan 48202, United States ; Xueqing Guo (1) ; Zhimeng Wu (1) . (1) Department<br />
<strong>of</strong> <strong>Chemistry</strong>, Wayne State University, Detroit Michigan 48202, United States<br />
Many surface proteins/glycoproteins are anchored onto the cell membrane via<br />
glycosylphosphatidylinositols (GPIs) – a class <strong>of</strong> complex glycolipids. GPIanchored<br />
proteins/glycoproteins play an important role in many biological<br />
processes. To study these processes, it is necessary to have access to<br />
homogeneous and structurally defined GPI-linked proteins, glycoproteins, and<br />
derivatives, which is currently difficult to achieve both biologically and chemically.<br />
To address the issue, we developed a practical method for GPI-linked<br />
protein/glycoprotein synthesis based on sortase-mediated site-specific ligation <strong>of</strong><br />
proteins/glycoproteins and chemically synthetic GPIs, as outlined in<br />
. In specific, sortase A (SrtA) <strong>of</strong> Staphylococcus aureus was used to couple GPIs<br />
and proteins/glycoproteins. SrtA recognizes a pentapeptide LPXTG near the<br />
substrate protein C-terminus, breaks the peptide bond between T and G, and<br />
then links the protein to a GPI analogue. The method has been employed to<br />
prepare a number <strong>of</strong> GPI-linked peptides, glycopeptides, and proteins and is<br />
expected to be generally applicable.
CARB 15<br />
Cancer relevant epitopes uncovered by synthetic mucin glycopeptides<br />
Shin-Ichiro Nishimura (1) , shin@glyco.sci.hokudai.ac.jp, N21 W11, Kita-ku,,<br />
Sapporo Hokkaido 0010021, Japan . (1) Hokkaido University, Sapporo Hokkaido<br />
0010021, Japan<br />
Robust compound library <strong>of</strong> synthetic MUC1 glycopeptides allowed for the first<br />
time rapid and precise identification <strong>of</strong> the specific epitope recognized by anti-KL-<br />
6 monoclonal antibody, a probe for detecting biomarker <strong>of</strong> interstitial pneumonia.<br />
We revealed that an essential epitope recognized by anti-KL-6 MAb is Pro-Asp-<br />
Thr-Arg-Pro-Ala-Pro in which Thr is modified by Neu5Aca2,3Galb1,3GalNAca.<br />
Anti-KL-6 MAb could not differentiate this structure from core 2-based<br />
glycopeptides involving this epitope and showed a similar binding affinity toward<br />
these compounds, indicating that branching at O-6 position <strong>of</strong> GalNAc does not<br />
influence the interaction <strong>of</strong> anti-KL-6 MAb with MUC1s involving an essential<br />
epitope. This is the reason why anti-KL-6 MAb <strong>of</strong>ten reacts with tumor-derived<br />
MUC1s as well as a biomarker <strong>of</strong> interstitial pneumonia, namely KL-6 originally<br />
discovered as a circulating pulmonary adenocarcinoma-associated antigen.<br />
Novel monoclonal antibodies obtained by this epitope reacted specifically with<br />
core 1-based structures and did not recognize MUC1s bearing core 2 type Oglycans.<br />
CARB 16<br />
<strong>Chemical</strong> biology <strong>of</strong> O-GlcNAc processing enzymes<br />
David J Vocadlo (1)(2) , dvocadlo@sfu.ca, 8888 University Drive, Burnaby British<br />
Columbia V5A 1S6, Canada ; Gideon J Davies (3) ; Tracey M Gloster (1) ; Julia E<br />
Heinonen (1) ; David Shen (2) ; Carlos Martinez-Fleites (3) ; Wesley F Zandberg (2) ;<br />
Yuan He (3) ; Thomas Clark (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, Simon Fraser<br />
University, Burnaby British Columbia V5A 1S6, Canada (2) Department <strong>of</strong><br />
Molecular Biology and Biochemistry, Simon Fraser University, Burnaby British<br />
Columbia V5A 1S6, Canada (3) Department <strong>of</strong> <strong>Chemistry</strong>, University <strong>of</strong> York,<br />
York, United Kingdom<br />
A common form <strong>of</strong> protein glycosylation in which serine and threonine residues <strong>of</strong><br />
nuclear and cytoplasmic proteins are post-translationally modified with O-linked<br />
2-acetamido-2-deoxy-β-D-glucopyranose residues (O-GlcNAc) is found in the<br />
nucleocytoplasm <strong>of</strong> multicellular eukaryotes. Unlike better known forms <strong>of</strong><br />
glycosylation occuring within the secretory pathway, O-GlcNAc is a dynamic<br />
modification that is turned over more rapidly than the proteins that it ornaments.<br />
Dysregulation <strong>of</strong> O-GlcNAc has been implicated in the etiology <strong>of</strong> various<br />
diseases. Two enzymes process O-GlcNAc; O-GlcNAc transferase (OGT)<br />
mediates installation <strong>of</strong> O-GlcNAc and O-GlcNAcase (OGA) catalyzes removal <strong>of</strong>
O-GlcNAc from proteins. Here we describe research into understanding the<br />
chemistry and biochemistry <strong>of</strong> these enzymes as well as how this knowledge has<br />
been used to develop chemical tools for manipulating O-GlcNAc levels in tissues.<br />
CARB 17<br />
Synthesis <strong>of</strong> a single molecule L-rhamnose-containing three component<br />
vaccine and evaluation <strong>of</strong> antigenicity in the presence <strong>of</strong> anti L-rhamnose<br />
antibodies<br />
Sourav Sarkar (1) , ssarkar2@UTNet.UToledo.Edu, 2801 W. Bancr<strong>of</strong>t St., Toledo<br />
OH 43606, United States ; Rommel S Talan (1) ; Steven A Lombardo (2) ; Katherine<br />
A Wall (2) ; Steven J Sucheck (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, The University <strong>of</strong><br />
Toledo, Toledo OH 43606, United States (2) Department <strong>of</strong> Medicinal and<br />
Biological <strong>Chemistry</strong>, The University <strong>of</strong> Toledo, Toledo OH 43606, United States<br />
We hypothesized that conjugation <strong>of</strong> L-rhamnose (Rha) to a carbohydrate<br />
antigen would enhance antigenicity <strong>of</strong> the antigen in mice possessing anti-Rha<br />
antibodies. We synthesized a vaccine containing the GalNAc-O-Thr (Tn) tumor<br />
specific antigen, a T-cell epitope (YAF) and a Rha moiety. Mice were immunized<br />
with Rha-ovalbumin (Rha-OVA). Anti-Rha antibody titers were >100 fold higher in<br />
groups <strong>of</strong> mice immunized with Rha-OVA than the control groups. Mice<br />
producing anti-Rha were challenged with Rha-YAF-Tn or YAF-Tn. Sera collected<br />
from the groups immunized with Rha-OVA and later challenged with Rha-YAF-Tn<br />
showed a two fold increase in anti-Tn titer at 1/100 serum dilution compared to<br />
mice not immunized with Rha-OVA. Proliferation <strong>of</strong> T-cells using cells primed<br />
with either Rha-YAF-Tn or YAF-Tn showed a 4-fold increase in the presence <strong>of</strong><br />
Rha antibodies. The results suggest that T-cells were presented with higher<br />
concentrations <strong>of</strong> Rha-YAF-Tn as a result <strong>of</strong> the presence <strong>of</strong> the anti-Rha<br />
antibodies.<br />
CARB 18<br />
Design <strong>of</strong> glycoprotein vaccines against influenza and other viral diseases<br />
Chi-Huey Wong (1)(2) , chwong@gate.sinica.edu.tw, 128 Academia Road, Section<br />
2, Nankang District, Taipei 115, Taiwan Republic <strong>of</strong> China . (1) Academia Sinica,<br />
Taipei 115, Taiwan Republic <strong>of</strong> China (2) The Scripps Research Institute, Taipei<br />
115, Taiwan Republic <strong>of</strong> China<br />
Protein glycosylation is the most complex post-translational process; more than<br />
90 percent <strong>of</strong> human proteins are glycosylated. The significance <strong>of</strong> glycosylation<br />
at the molecular level is however not well understood, and as such the pace for<br />
the development <strong>of</strong> carbohydrate-based drug discovery and diagnosis is<br />
relatively slow. It is thus important to develop new tools to study the effect <strong>of</strong><br />
glycosylation on the structure and function <strong>of</strong> proteins and other biologically
active molecules. This lecture will focus on the development <strong>of</strong> new methods for<br />
the synthesis <strong>of</strong> homogenious glycoproteins with well defined glycan structure,<br />
glycoarrays for the high-throughput analysis <strong>of</strong> protein-glycan interaction and<br />
design <strong>of</strong> click-induced fluorescent probes for use to identify new cancer<br />
biomarkers for diagnosis and drug discovery. New glycoprotein vaccines have<br />
been designed and developed to tackle the problems <strong>of</strong> flu and breast cancer.<br />
CARB 19<br />
Modular synthesis <strong>of</strong> heparan sulfate oligosaccharides for array<br />
development<br />
Geert-Jan Boons (1) , gjboons@ccrc.uga.edu, 315 Riverbend Rd, Athens GA<br />
30606, United States . (1) Complex <strong>Carbohydrate</strong> Research Center, University <strong>of</strong><br />
Georgia, Athens Georgia 30606, United States<br />
Although hundreds <strong>of</strong> heparan sulfate binding proteins have been identified, and<br />
implicated in a myriad <strong>of</strong> physiological and pathological processes, very little<br />
information is known about ligand requirements for binding and mediating<br />
biological activities by these proteins. This difficulty results from a lack <strong>of</strong><br />
technology for establishing structure-activity-relationships, which in turn is due to<br />
the structural complexity <strong>of</strong> natural HS and difficulties <strong>of</strong> preparing well-defined<br />
HS-oligosaccharides. To address this deficiency, we have developed a modular<br />
approach for the parallel combinatorial synthesis <strong>of</strong> HS oligosaccharides that<br />
utilizes a relatively small number <strong>of</strong> selectively protected disaccharide building<br />
blocks, which can easily be converted into glycosyl donors and acceptors. The<br />
modular building blocks have been used for the preparation <strong>of</strong> a library <strong>of</strong><br />
oligosaccharides, which has been employed to probe ligand requirements <strong>of</strong><br />
several HS-binding proteins. To facilitate high through put screening, spacer<br />
modified HS-oligosaccharides have been employed for microarray fabrication.<br />
CARB 20<br />
New targets for antibiotics<br />
Suzanne Walker (1) , suzanne_walker@hms.harvard.edu, 200 Longwood Ave,<br />
Boston MA 02115, United States . (1) Department <strong>of</strong> Microbiology and Molecular<br />
Genetics, Harvard Medical School, Boston MA 02115, United States<br />
Wall teichoic acids (WTAs) are anionic carbohydrate polymers that are covalently<br />
attached to the peptidoglycan layers <strong>of</strong> many Gram positive organisms, including<br />
S. aureus. They comprise 50% <strong>of</strong> the cell wall mass and have a major influence<br />
on cell envelope integrity. They are suggested targets for antibiotics since<br />
deleting certain genes in the pathway results in a loss <strong>of</strong> virulence while deleting<br />
others is lethal. We developed a pathway-specific, cell based screen to discovery<br />
inhibitors <strong>of</strong> WTA biosynthesis and have identified a potent WTA-active antibiotic.
We will describe studies on the mechanism <strong>of</strong> action <strong>of</strong> this compound and on<br />
the cellular effects <strong>of</strong> blocking wall teichoic acid biosynthesis. Prospects for the<br />
use <strong>of</strong> WTA inhibitors as antibiotics will be discussed.<br />
CARB 21<br />
Challenges and opportunities in natural product glycosylation<br />
Jon S Thorson (1) , jsthorson@pharmacy.wisc.edu, 777 Highland Avenue,<br />
Madison WI 53705-2222, United States . (1) School <strong>of</strong> Pharmacy, University <strong>of</strong><br />
Wisconsin, United States<br />
In nature, the attachment <strong>of</strong> sugars to small molecules is<br />
<strong>of</strong>ten employed to mediate targeting, mechanism <strong>of</strong> action, and/or<br />
pharmacology. As an alternative to<br />
pathway engineering or classical chemical glycosylation strategies, we report<br />
merging three promiscuous enzymes (a sugar kinase - 'E1', a<br />
nucleotidylyltransferase - 'E2', and a glycosyltransferase – 'E3')<br />
with upstream synthetic chemistry and downstream chemoselective ligation<br />
provides a powerful method to diversify, via the attachment <strong>of</strong> variant sugars<br />
('glycorandomize'), complex natural products.<br />
The optimization <strong>of</strong> E1-E3 using enzyme evolution<br />
and structure-based enzyme engineering, the glycorandomization pro<strong>of</strong> <strong>of</strong><br />
concept, the recent advances based upon the reversibility <strong>of</strong><br />
glycosyltransferase-catalyzed reactions, the evolution <strong>of</strong> 'flexible'<br />
glycosyltransferases and the potential for in<br />
vivo glycosylation hosts will be highlighted. In addition, recent developments<br />
pertaining<br />
to our development <strong>of</strong> a complementary chemical method<br />
('neoglycorandomization')<br />
will be discussed. The impact <strong>of</strong> the<br />
(neo)glycorandomization upon the activity <strong>of</strong> selected small molecule-based<br />
drugs and/or understanding sugar transport may also be presented.<br />
CARB 22<br />
Glycomimetic and non-carbohydrate inhibitors as probes <strong>of</strong> the lectin DC-<br />
SIGN<br />
Laura L Kiessling (1) , kiessling@chem.wisc.edu, 1101 University Avenue,<br />
Madison WI 53706, United States . (1) Departments <strong>of</strong> <strong>Chemistry</strong> and<br />
Biochemistry, University <strong>of</strong> Wisconsin-Madison, Madison WI, United States<br />
<strong>Carbohydrate</strong>s act through carbohydrate-binding proteins (lectins) to govern<br />
numerous cellular processes, including cell adhesion, recognition, and signaling.<br />
Despite the importance <strong>of</strong> lectins, inhibitors that can be used to interrogate or
mitigate their function are rare. In principle, carbohydrates themselves can serve<br />
as probes, but they have several liabilities, including low affinity and relaxed<br />
specificity. We have used high throughput screening and chemical synthesis to<br />
identify potent inhibitors <strong>of</strong> the lectin DC-SIGN. DC-SIGN is a C-type lectin found<br />
on the surface <strong>of</strong> dendritic cells that mediates antigen uptake and dendritic cell–T<br />
cell interactions. Ironically, this immune system lectin can be co-opted to facilitate<br />
the dissemination <strong>of</strong> pathogens, including HIV, dengue virus, and Mycobacterium<br />
tuberculosis. The identification <strong>of</strong> DC-SIGN inhibitors provides the means to<br />
dissect the function <strong>of</strong> this lectin and its involvement in a wide range <strong>of</strong> infectious<br />
diseases.<br />
CARB 23<br />
Toxicity and functional assessment using polysaccharide-based magnetic<br />
iron oxide nanoparticles for cell labeling in vivo and in vitro<br />
Yoshitaka Miyamoto (1) , myoshi1230@hotmail.com, Higashi-ku, Nagoya 461-<br />
0047, Japan ; Yumie Koshidaka (1) ; Hiroaki Saito (2) ; Yukimasa Kagami (3) ;<br />
Katsutoshi Murase (2) ; Noritada Kaji (3) ; Hiroshi Yukawa (1) ; Hir<strong>of</strong>umi Noguchi (4) ;<br />
Hisashi Iwata (5) ; Yoshinobu Baba (3) ; Shuji Hayashi (1) . (1) Department <strong>of</strong><br />
Advanced Medicine in Biotechnology and Robotics, Nagoya University Graduate<br />
School <strong>of</strong> Medicine, Higashi-ku Nagoya 461-0047, Japan (2) Nagoya Research<br />
Laboratory, MEITO Sangyo Co., Ltd., 25-5, Kaechi, Nishibiwajima Kiyosu 452-<br />
0067, Japan (3) Department <strong>of</strong> Applied <strong>Chemistry</strong>, Nagoya University Graduate<br />
School <strong>of</strong> Engineering, Furo-cho, Chikusa-ku Nagoya 464-8603, Japan (4)<br />
Baylor Institute for Immunology Research, Baylor Research Institute, 3434 Live<br />
Oak St., Dallas TX 75204, United States (5) Department <strong>of</strong> Biomedical Sciences,<br />
Chubu University College <strong>of</strong> Life and Health Sciences, 1200 Matsumoto-cho,<br />
Kasugai Aichi 487-8501, Japan<br />
It is desireble to develop less-invasive medical treatment and diagnostics for the<br />
patient in breakthrough <strong>of</strong> medical technology. Especially, contrast agents for<br />
magnetic resonance imaging (MRI) have been widely used to diagnose blood<br />
vessel disorder, internal structure <strong>of</strong> organs. Recently, we developed a novel<br />
contrast agent for MRI using polysaccharide-based magnetic iron oxide<br />
nanoparticles. The agent was transduced into each cell efficiently. Additionally,<br />
we reported that the effective imaging <strong>of</strong> transplanted cells was the efficienty<br />
labeled using the contrast agent. The purpose <strong>of</strong> this study is to evaluate the<br />
effect <strong>of</strong> the contrast agent on cell functions in vivo and in vitro. In this<br />
experiment, the contrast agent was added to human hepatocellular liver<br />
carcinoma cell line (HepG2) and its effect on cell functions was evaluated. As a<br />
result, the dependence <strong>of</strong> the amount <strong>of</strong> a contrast agent on liver functions was<br />
investigated using different contrast agents.<br />
CARB 24
Glycodendrimers for the mediation <strong>of</strong> cancer cellular aggregation<br />
processes<br />
Mary J Cloninger (1) , mcloninger@chemistry.montana.edu, 103 <strong>Chemistry</strong> and<br />
Biochemistry Building, Bozeman Montana 59717, United States ; Mark L.<br />
Wolfenden (1) ; Julie J Sprenger (1) ; Michael P Capp (1) ; Pratima Nangia-Makker (2) ;<br />
Avraham Raz (2) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and Biochemistry, Montana State<br />
University, Bozeman Montana 59717, United States (2) Department <strong>of</strong><br />
Pathology, School <strong>of</strong> Medicine, Wayne State University, Detroit MI 48201, United<br />
States<br />
Galectin-3 is a galactoside-binding protein that is commonly up or down<br />
regulated in different cancers and is implicated in tumor formation and<br />
proliferation, apoptosis, angiogenesis, and B cell activation. In particular,<br />
galectin-3 is thought to be involved in the aggregation processes that lead to<br />
tumor proliferation, making this lectin an interesting target for our multivalent<br />
carbohydrate functionalized poly(amidoamine) (PAMAM) dendrimers.<br />
Capitalizing on the differences in the monovalent binding interaction between<br />
galactose, lactose, N-acetylgalactosamine, and N-acetyllactosamine, and also on<br />
the range <strong>of</strong> generations that are readily available for PAMAM dendrimers, a<br />
library <strong>of</strong> glycodendrimers was prepared. This series <strong>of</strong> glycodendrimers includes<br />
compounds having a large range <strong>of</strong> binding capabilities with galectin-3 and<br />
enables attenuation <strong>of</strong> galectin-3 mediated biological processes. Here, the<br />
binding interactions <strong>of</strong> these glycodendrimers with galectin-3 and the effect that<br />
glycodendrimers have on cancer cellular aggregation processes are presented.<br />
CARB 25<br />
Glyconanoparticles: Multibi<strong>of</strong>unctional nanomaterials for biomedical<br />
applications<br />
Soledad Penades (1) , spenades@cicbiomagune.es, Paseo Miramon, 182, San<br />
Sebastian Guipuzcoa 20009, Spain . (1) Bi<strong>of</strong>unctional Nanomaterials Unit, CIC<br />
biomaGUNE, San Sebastian Guipuzcoa 20009, Spain<br />
Our laboratory pioneered the development <strong>of</strong> a new technology<br />
(Glyconanotechnology) for tailoring - in a simple and versatile way – bi<strong>of</strong>unctional<br />
gold nanoclusters (glyconanoparticles, GPNs). GNPs present some<br />
advantages over other previously prepared colloids as: 1) easy preparation and<br />
purification; 2) exceptional small core size and narrow distribution sizes; 3)<br />
control over ligands number and nanoparticles size; 4) water solubility; 5) high<br />
storage stability without flocculation; and 6) singular physical properties.<br />
GNPs with biologically significant carbohydrates and with differing carbohydrate<br />
density were prepared to study and intervene in carbohydrate-mediate cell<br />
adhesion processes. The methodology includes also the preparation <strong>of</strong> hybrid
GNPs incorporating carbohydrates and other molecules such as fluorescent<br />
probes, biotin as well as biological molecules such as peptides, DNA and RNA<br />
providing with the possibility to create artificial “nanocells”.<br />
The manipulation <strong>of</strong> the metallic cluster to obtain luminescent glyco-quantum<br />
dots (semiconductors) and magnetic glyco-nanoparticles for application in<br />
cellular labeling and imaging by magnetic resonance (MRI), is comprised within<br />
the potential <strong>of</strong> this technology. Examples <strong>of</strong> gold GNPs as anti-adhesion agents<br />
and as magnetic probes in cellular labeling and tracking by magnetic resonance<br />
imaging (MRI) will be highlighted.<br />
CARB 26<br />
New glycopeptide-based nanoparticle constructions for anticancer therapy<br />
Joseph John Barchi (1) , barchi@helix.nih.gov, 376 Boylse Street, PO Box B,<br />
Frederick MD 21702, United States ; Ray Brinas (2) ; Andreas Sundgren (1) ;<br />
Padmini Sahoo (3) ; Amy Houghten (3) ; Susan Morey (3) ; Michael Sanford (2) ; Howard<br />
Young (2) . (1) <strong>Chemical</strong> Biology, National Cancer Institute at Frederick, Frederick<br />
MD 21702, United States (2) Experimental Immunology, National Cancer<br />
Institute at Frederick, Frederick MD 21702, United States (3) Biotechnical and<br />
Clinical Laboratory Sciences, University <strong>of</strong> Buffalo, Buffalo NY 14214, United<br />
States<br />
The surface <strong>of</strong> mammalian cells is replete with carbohydrates that are presented<br />
in a wide range <strong>of</strong> structural subtypes and are involved in a wealth <strong>of</strong> biologically<br />
important functions. Transformation <strong>of</strong> normal cells to a neoplastic phenotype<br />
involves aberrant gene expression resulting in the presence <strong>of</strong> very different<br />
glycans displayed on the tumor cell surface. Many <strong>of</strong> these are recognized and<br />
targeted by the immune system and are thus referred to as tumor-associated<br />
carbohydrate antigens (TACA). Various TACA's have been employed in anti<br />
tumor vaccines with mostly disappointing results due to immunological tolerance<br />
and lack <strong>of</strong> a cell mediated response. We propose here a novel strategy for the<br />
preparation <strong>of</strong> fully synthetic immunogens based on a nanoscale inorganic<br />
platform that displays tumor glycopeptide antigens in a multivalent fashion. We<br />
have linked these to multicomponent gold nanoparticles as “stand alone”<br />
immunogens where an appropriate nanoconstruction is used in place <strong>of</strong><br />
immunostimulating proteins. The talk will concentrate on the development <strong>of</strong><br />
these and other particles as vaccine constructs and novel entities that elicit<br />
tailored cytokine responses when they interact with antigen presenting cells.<br />
CARB 27<br />
Functionalized catanionic surfactant vesicles: A new approach to<br />
carbohydrate vaccines
Philip DeShong (1) , deshong@umd.edu, University <strong>of</strong> Maryland, Department <strong>of</strong><br />
<strong>Chemistry</strong> and Biochemistry, College Park MD 20742, United States ; Juhee<br />
Park (1)(2) ; Lenea Rader (1) ; Glen Thomas (1) ; Douglas English (2)(3) ; Lindsey<br />
Zimmerman (4) ; Daniel C. Stein (4) . (1) Department <strong>of</strong> <strong>Chemistry</strong> & Biochemistry,<br />
University <strong>of</strong> Maryland, College Park MD 20742, United States (2) SD<br />
Nanosciences, Inc., College Park MD 20742, United States (3) Department <strong>of</strong><br />
<strong>Chemistry</strong>, Wichita State University, College Park MD 20742, United States (4)<br />
Department <strong>of</strong> Cell Biology & Molecular Genetics, University <strong>of</strong> Maryland,<br />
College Park MD 20742, United States<br />
Vaccines against Gram-negative pathogens (e.g. E. coli, F. tularensis, N.<br />
meningitidis, P. aeruginosa) are difficult to produce because their glycosylated<br />
cell surfaces (i.e. lipolysaccharides) are poorly immunogenic and fail to elicit a<br />
helper T-cell mediated immune response. Catanionic surfactant vesicles are<br />
unilamellar vesicles that form spontaneously from a mixture <strong>of</strong> cationic and<br />
anionic surfactants in water. The size and surface charge <strong>of</strong> the vesicle can be<br />
controlled by choice <strong>of</strong> surfactants, and the external leaflet <strong>of</strong> the vesicle can be<br />
decorated with non-ionic surfactants in concentrations as high as 20 mole<br />
percent without affecting the stability <strong>of</strong> the vesicle. We incorporated a mixture <strong>of</strong><br />
peptides designed to interact with T-cells and LOS-derived glycoconjugates into<br />
catanionic vesicles. The resulting functionalized vesicle was capable <strong>of</strong> inducing<br />
a strong IgG response in mice. The synthesis and characterization <strong>of</strong> the vesicles<br />
will be discussed and biological evaluation <strong>of</strong> this approach to carbohydrate<br />
vaccines will be presented.<br />
CARB 28<br />
Novel synthesis <strong>of</strong> 2'-deoxynucleoside 5'-triphosphates without nucleoside<br />
protection and DNA polymerase recognition<br />
Julianne M. Caton-Williams (1) , jcatonwilliams1@gsu.edu, Room 540 General<br />
Classroom Building, 38 Peachtree Center Ave., Atlanta Georgia 30303-3083,<br />
United States ; Matthew R. Smith (1) ; Zhen Huang (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong><br />
and Department <strong>of</strong> Biology, Georgia State University, Atlanta Georgia 30303,<br />
United States<br />
Because nucleosides comprise amino and multi-hydroxyl groups, the synthesis<br />
<strong>of</strong> the nucleoside 5'-triphosphates requires protection and deprotection <strong>of</strong> these<br />
groups, which are achieved in multiple synthetic steps. In order to simplify the<br />
triphosphate synthesis and avoid the protection and deprotection reactions, we<br />
have developed a convenient strategy to synthesize 2'-deoxynucleoside 5'triphosphates<br />
(dNTPs) without nucleoside protection. The facile synthesis is<br />
achieved by generating an in situ, selective phosphorylating reagent that reacts<br />
selectively with the 5'-hydroxyl group and finally generates the triphosphates in<br />
one-pot. The synthesized triphosphates are <strong>of</strong> high quality and can be effectively<br />
incorporated into DNAs by DNA polymerase. The developed synthetic conditions
are mild, and this strategy is also useful in one-pot synthesis <strong>of</strong> modified<br />
triphosphates.<br />
CARB 29<br />
Activation <strong>of</strong> retinoic acid<br />
inducible gene (RIG-I) by short oligonucleotides<br />
Radhakrishnan Iyer (1) , hcato@alnylam.com, 113 Cedar Street, Milford MA,<br />
United States ; John Coughlin (1) ; Seetharamaiyer Padmanabhan (1) ; Brent<br />
Korba (2) ; Sua Myong (3) . (1) Spring Bank Pharmaceuticals, United States (2)<br />
Georgetown University, United States (3) University <strong>of</strong> Illinois- Urbana-<br />
Champaign Department, United States<br />
Retinoic acid<br />
inducible gene (RIG-I), a host cellular protein, acts as a<br />
“sensor” <strong>of</strong> viral RNA. Activation <strong>of</strong> RIG-I results in<br />
Interferon (IFN) expression and induction <strong>of</strong> antiviral effects. Using a novel<br />
translocation assay <strong>of</strong> RIG-I on a<br />
dsRNA template, we discovered that short oligonucleotides (SO) cause activation<br />
<strong>of</strong> RIG-I. The RIG-I<br />
actives had specific structural and stereochemical attributes. The RIG-I<br />
active analog SB 40 had potent antiviral activity against HBV (EC50,<br />
0.5 to 1 micromolar in HepG2.2.15 cell lines) and was synergistic with 3TC and<br />
Adefovir. SB 40 showed potent<br />
antiviral activity in the HBV transgenic mouse model with an EC50 <<br />
1 mg/kg. A related analog SB 44 had EC50<br />
(<strong>of</strong> 1~2 micromolar) against HCV (genotypes 1a, 1b) with synergistic antiviral<br />
activity with other anti-HCV drugs. The stimulation <strong>of</strong> RIG-I pathway<br />
by SO could be exploited for multiple therapeutic applications including<br />
discovery <strong>of</strong> orally bioavailable broad-spectrum antiviral agents.<br />
CARB 30<br />
Solid-phase synthesis <strong>of</strong> 5'-di- and tri-phosphates and<br />
their modified analogs <strong>of</strong> DNA, RNA and chemically modified<br />
oligonucleotides<br />
Ivan Zlatev (1) , izlatev@alnylam.com, 300 3rd St, 3rd Floor, Cambridge MA,<br />
United States ; Thomas Lavergne (2) ; Sudhakar Takkellapati (1) ; Rajenda<br />
Pandey (1) ; Yupeng Fan (1) ; Marija Prhavc (1) ; Kathy Mills (1) ; G. Rajeev<br />
Kallanthottathil (1) ; Françoise Debart (2) ; Jean-Jacques Vasseur (2) ; François<br />
Morvana (2) ; Muthiah Manoharan (1) . (1) Alnylam Pharmaceuticals, United States<br />
(2) Institut des Biomolécules Max Mousseron, France
A<br />
robust, reproducible,<br />
scalable, and automated method<br />
for the solid-phase synthesis <strong>of</strong> 5'-di- and tri-phosphates <strong>of</strong> DNA, RNA, and<br />
their chemically modified analogs that employs 5'-H-phosphonate intermediates<br />
is described. 5'-Triphosphates <strong>of</strong><br />
oligonucleotides <strong>of</strong> varying lengths and sequences containing different<br />
5'-terminal nucleotides with and without internal sugar and/or backbone<br />
modifications (2'-O-Me, 2'-F, 2'-O-MOE, LNA, phosphorothioate)<br />
were efficiently prepared as crude products or further purified by HPLC.<br />
The synthetic scheme involved the introduction <strong>of</strong> a 5'-H-phosphonate<br />
monoester onto a solid-supported oligonucleotide; the 5'-H-phosphonate<br />
monoester was further transformed into either a di-<br />
or tri-phosphate. Oligonucleotide 5'-polyphosphates were typically obtained in<br />
high yields and acceptable purity. The preparation method was efficiently<br />
adapted to a fully automated synthesis cycle using a standard oligonucleotide<br />
synthesizer.<br />
CARB 31<br />
Drug discovery harnessing RNA interference<br />
Muthiah Manoharan (1) , mmanoharan@alnylam.com, 300 3rd St, 3rd Floor,<br />
Cambridge MA, Qatar . (1) Alnylam Pharmaceuticals, United States<br />
RNA interference (RNAi) is a powerful biological<br />
process for specific silencing <strong>of</strong> mRNAs in diverse eukaryotic cells. By using<br />
synthetic short interfering RNAs (siRNAs) this process can be utilized to<br />
develop drugs targeting any given gene, including the “undruggable targets”.<br />
Desirable<br />
"drug-like" properties can be imparted to siRNAs by introducing<br />
chemically modified nucleosides and other synthetic building blocks. siRNAs<br />
containing chemical modifications show enhanced resistance towards nuclease<br />
degradation,<br />
less immune stimulation, and reduced "<strong>of</strong>f-target" effects compared to<br />
unmodified siRNAs. To achieve in vivo<br />
delivery, certain chemical conjugates and novel liposomal formulations are<br />
being investigated. A summary <strong>of</strong> this drug discovery paradigm and<br />
progress in the development <strong>of</strong> several new drug compounds will be presented<br />
CARB 32<br />
Parallel high throughput synthesis <strong>of</strong> chemically modified 21-27mer<br />
siRNA sequences
Jason Costigan (1) , jcostigan@alnylam.com, 300 3rd St, 3rd Floor, Cambridge<br />
MA, United States ; Sarfraz Shaikh (1) ; Keri Dufault (1) ; Jack de Groot (1) ; Satya<br />
Kuchimanchi (1) ; Rachel Meyers (1) . (1) Alnylam Pharamaceuticals, United States<br />
Methods have been developed for high throughput synthesis <strong>of</strong><br />
21 to 27mer siRNA sequences containing different chemical modifications. We<br />
have developed procedures that enable rapid generation <strong>of</strong> modified RNA single<br />
strands and duplexes using a MerMade192 synthesis platform. The purpose <strong>of</strong><br />
this work is to generate high<br />
quality duplexes for in vitro screening experiments and bio-analytical assays.<br />
Sequences that contain different chemical<br />
modifications, including 2'OMe, 2'F, 5' phosphate and 3' cholesterol have been<br />
successfully synthesized and fully characterized by LC-MS and analytical ion<br />
exchange methods. Cleavage and deprotection methods have been<br />
optimized to minimize formation <strong>of</strong> impurities. Development <strong>of</strong> high throughput<br />
purification processes and annealing methods that complement high throughput<br />
synthesis productivity will be discussed.<br />
CARB 33<br />
New developments in the synthesis <strong>of</strong> oligoribonucleotides: Use <strong>of</strong><br />
dimer/trimer blocks in combination with an “ionic tag” soluble support<br />
Matthew R Hassler (1) , matthew.hassler@mail.mcgill.ca, 801 Sherbrooke St. W,<br />
Montreal Quebec H3A 2K6, Canada ; Nandyala Mallikarjuna Reddy (1) ; Tak-Hang<br />
Chan (1) ; Masad J Damha (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, McGill University,<br />
Montreal Quebec H3A 2K6, Canada<br />
The reality <strong>of</strong> mainstream RNA based therapeutics is quickly approaching and<br />
the demand for these compounds may soon exceed the capabilities <strong>of</strong> the<br />
manufacturers. Large-scale synthesis <strong>of</strong> short-interfering RNA (siRNA) is<br />
generally carried out via iterative step-wise solid phase synthesis. A major<br />
drawback <strong>of</strong> the solution-phase alternative is the rigorous purification required<br />
between each step, which can be costly and time consuming. We have<br />
demonstrated that by using an “ionic tag” soluble support we are able to<br />
overcome some <strong>of</strong> the limitations <strong>of</strong> solution-phase synthesis. This allows for<br />
selective precipitation <strong>of</strong> the growing RNA over all other reagents used in the<br />
oligonucleotide synthesis cycle. Herein, we will present our recent developments<br />
in this area. Additionally, we will describe the synthesis <strong>of</strong> di- and tri- nucleotide<br />
amidite building blocks for large scale RNA solution phase synthesis. siRNAs<br />
synthesized by our newly developed method will be compared to traditional solidphase<br />
synthesized siRNAs for purity and biological activity.<br />
CARB 34<br />
Advanced process <strong>of</strong> RNA synthesis
Anuj Mohan (1) , amohan@chemgenes.com, 33 Industrial Way, Wilmington MA<br />
01887, United States . (1) Chemgenes, United States<br />
We have developed a process <strong>of</strong> RNA synthesis that utilizes 3'- DMT -5'- CED<br />
phosphoramidites leading to step-wise coupling efficiency > 99% in the reverse<br />
RNA synthesis (5 ->3') resulting in high purity short and long RNA sequences.<br />
These new monomers have been found to be distinctly superior to the standard<br />
3'- CED -2'-O-TBDMS phosphoramidites used in conventional RNA synthesis (3'<br />
-> 5'). The monomers provide method for synthesis <strong>of</strong> clean RNA with wide<br />
variety <strong>of</strong> modifications or labels at 3'- end <strong>of</strong> oligonucleotide. The mechanism <strong>of</strong><br />
the novel process will be discussed.<br />
CARB 35<br />
Novel method for the confirmation <strong>of</strong> siRNA sequence by<br />
LC-MS/MS<br />
Gary Lavine (1) , glavine@alnylam.com, 300 3rd St, 3rd Floor, Cambridge MA,<br />
United States ; Matthias Kretschmer (1) ; James McArdle (2) ; Satya Kuchimanchi (1) ;<br />
Veeragu Murugaiah (1) ; Muthiah Manoharan (1) . (1) Alnylam Pharmaceuticals,<br />
United States (2) McArdle & Associates, LLC, United States<br />
We have developed a new procedure for confirmation <strong>of</strong> the<br />
sequence <strong>of</strong> chemically modified synthetic oligonucleotides by tandem mass<br />
spectrometry fragmentation. The method is highly reliable for the<br />
interpretation <strong>of</strong> high-resolution Q-TOF MS/MS data. Deconvolution <strong>of</strong> the<br />
MS/MS data provided<br />
simplified peak assignments were used to match expected fragment ions. Data<br />
from the MS/MS experiments <strong>of</strong> three<br />
different charge states were combined; evaluation <strong>of</strong> these complementary data<br />
sets provided more complete coverage. To<br />
verify the assigned structure, the sequence confirmation was challenged by<br />
comparison <strong>of</strong> the quality <strong>of</strong> fit to computer-generated sequence<br />
perturbations. Through this novel<br />
perturbation method we have shown that the sequence <strong>of</strong> synthetic<br />
oligonucleotides can be confirmed using MS/MS data from a Q-TOF instrument.<br />
CARB 36<br />
Evaluation <strong>of</strong> Canonical vs.<br />
Dicer-substrate siRNAs in vitro and in vivo<br />
Don Foster (1) ; Satya Kuchimanchi (1) ; Greg Hinkle (1) ; William Cantley (1) ; Rick<br />
Duncan (1) ; Geff Cole (1) ; Chris Sherill (1) ; Kathy Mills (1) ; Mara Broberg (1) ; Jeff<br />
Rollins (1) ; Klaus Charisse (1) ; Muthiah Manoharan (1) ; Sayda Elbashir (1) . (1) Alnylam<br />
Pharmaceuticals, United States
Canonical<br />
siRNAs are duplexes formed by two 21-mers; 19 bases are paired and two<br />
nucleotides overhang on each strand (Elbashir SM et al., 2001, RNA interference<br />
is mediated by 21- and 22-nucleotide RNAs. Genes<br />
Dev. 15:.188-200; Elbashir SM, Harborth J, Lendeckel W,<br />
Yalcin A, Weber K and Tuschl T., 2001, Duplexes <strong>of</strong> 21-nucleotide RNAs<br />
mediate<br />
RNA interference in cultured mammalian cells. Nature 411:<br />
494–498). Dicer-substrate siRNAs (DsiRNA) are 25/27-mer duplexes<br />
from which 21-mer siRNAs are generated by the action <strong>of</strong> Dicer in situ (Kim DH,<br />
Behlke MA, Rose SD, Chang MS, Choi S and Rossi JJ. 2005,<br />
Synthetic dsRNA Dicer substrates enhance RNAi potency and efficacy. Nat.<br />
Biotechnol. 23: 222–226).We<br />
conducted comprehensive studies in vitro<br />
and in vivo to compare canonical<br />
siRNA and DsiRNA. We chose two well-characterized genes as targets and<br />
designed<br />
over 300 compounds, representing both types <strong>of</strong> siRNAs, to hybridize to these<br />
mRNAs. We will present data from this comparative study regarding silencing<br />
efficacy, duration <strong>of</strong> silencing, and other key parameters such as nuclease<br />
stability and cytokine induction.<br />
CARB 37<br />
Modulation <strong>of</strong> thermal stability can enhance the potency <strong>of</strong> siRNA<br />
Haripriya Addepalli (1) , hAddepalli@alnylam.com, 300 3rd St, 3rd Floor,<br />
Cambridge MA, United States ; Meena Meena (1) ; Chang G. Peng (1) ; Gang<br />
Wang (1) ; Yupeng Fan (1) ; Klaus Charisse (1) ; K. Narayanannair Jayaprakash (1) ; G.<br />
Rajeev Kallanthottathil (1) ; Rajendra Pandey (1) ; Gary Lavine (1) ; Ligang Zhang (1) ;<br />
Kerstin Jahn-H<strong>of</strong>mann (2) ; Philipp Hadwiger (2) ; Muthiah Manoharan (1) ; Martni<br />
Maier (1) . (1) Alnylam Pharmaceuticals, United States (2) Alnylam Europe,<br />
Germany<br />
During RISC assembly, the guide (or antisense) strand<br />
must separate from its complementary passenger (or sense) strand to generate<br />
the active RISC complex. Although this process is facilitated through sense<br />
strand cleavage, there is evidence for an alternate mechanism, in which the<br />
strands are dissociated without cleavage <strong>of</strong> the sense strand. Here we show that<br />
the potency <strong>of</strong> siRNA can be improved by modulating the thermodynamic stability<br />
<strong>of</strong> the duplex formed between sense and antisense strands. We found that<br />
placement <strong>of</strong> thermally destabilizing modifications, such as a non-canonical<br />
bases like 2,4-difluorotoluene, or single base pair mismatches in the central<br />
region <strong>of</strong> the sense strand (nucleotides 9 to 12) significantly improved the<br />
potency <strong>of</strong> a well-validated siRNA targeting firefly luciferase. For this<br />
particular siRNA, the strongest correlation between the decrease in thermal
stability and the increase in potency was found at position 10. Controls with a<br />
stabilized sugar-phosphate backbone indicated that enzymatic cleavage <strong>of</strong> the<br />
sense strand prior to strand dissociation was not required for the observed<br />
silencing activity.<br />
CARB 38<br />
<strong>Carbohydrate</strong> conjugation to<br />
siRNA for tissue-specific delivery<br />
G. Rajeev Kallanthottathil (1) , rajeevk@alnylam.com, 300 3rd St, 3rd Floor,<br />
Cambridge MA, United States ; K. Narayanannair Jayaprakash (1) ; Maria Frank-<br />
Kamenetsky (1) ; Gang Wang (1) ; Tianlei Lei (1) ; Mariano Severgnini (1) ; William<br />
Querbes (1) ; Jim Butler (1) ; Alfica Sehgal (1) ; Tomoko Nakayama (1) ; Klaus Charrise (1) ;<br />
Martin Maier (1) ; Kevin Fitzgerald (1) ; Muthiah Manoharan (1) . (1) Alnylam<br />
Pharmaceuticals, United States<br />
Systemic administration <strong>of</strong><br />
lipophile-conjugated siRNA mediates uptake into cells and causes gene silencing<br />
in vivo (Nature Biotech., 2007, 25, 1149; Nature, 2004, 432, 173). To further<br />
improve<br />
tissue-specific delivery <strong>of</strong> siRNAs, we conjugated carbohydrate ligands to siRNA<br />
with and without lipophilic moieties. These ligands bind specifically to the<br />
asialoglycoprotein receptor (ASGPR). ASGPR is a transmembrane glycoprotein<br />
<strong>of</strong><br />
42 kDa that mediates binding, internalization, and degradation <strong>of</strong> extracellular<br />
glycoproteins with exposed terminal galactose residues. The receptor is highly<br />
expressed on the surface <strong>of</strong> liver hepatocytes; it is present on the sinusoidal<br />
and lateral plasma membranes. Among various galactose analogues tested for<br />
binding<br />
affinity, a triantennary N-acetylgalactosamine<br />
(GalNAc) cluster had nanomolar binding affinity to the receptor. We will<br />
describe synthesis <strong>of</strong> multivalent GalNAc-siRNA conjugates for<br />
hepatocyte-specific delivery and in vitro<br />
receptor binding, uptake, and gene silencing.<br />
CARB 39<br />
Sugar and proteins: Applications <strong>of</strong> bioconjugates<br />
Benjamin G. Davis (1) , Ben.Davis@chem.ox.ac.uk, <strong>Chemistry</strong> Research<br />
Laboratory, Oxford Oxfordshire OX1 3TA, United Kingdom . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, University <strong>of</strong> Oxford, Oxford OX1 3TA, United Kingdom<br />
Sugars and Post-Translational Modifications are critical biological markers that<br />
modulate the properties <strong>of</strong> proteins. Our work studies the interplay <strong>of</strong> proteins,
sugars and modifications<br />
and their utility.<br />
(i) Synthetic Biology's development at the start <strong>of</strong> this century may be compared<br />
with Synthetic Organic <strong>Chemistry</strong>'s expansion at the start <strong>of</strong> the last; after<br />
decades <strong>of</strong> isolation, identification, analysis and functional confirmation the future<br />
logical and free-ranging redesign <strong>of</strong> biomacromolecules <strong>of</strong>fers tantalizing<br />
opportunities. New methods are required: despite 80-years-worth <strong>of</strong> non-specific,<br />
chemical modification <strong>of</strong> proteins, precise methods in protein chemistry remain<br />
rare. The development <strong>of</strong> efficient, complete, chemo- and regio-selective<br />
methods, applied in benign aqueous systems to redesign the structure and<br />
function <strong>of</strong> bioconjugates (proteins, polymers, nanoparticles, viruses, nanotubes)<br />
will be presented.<br />
(ii) Bioconjugate Applications: These conjugates find application in • drug delivery<br />
• nanomolar inhibitors <strong>of</strong> bacterial interactions • gene delivery vehicles • probes<br />
<strong>of</strong> in vivo function • non-invasive presymptopmatic disease diagnosis • localized<br />
delivery <strong>of</strong> unprecedented radio-dose.<br />
[1] B.G.Davis, Chem. Rev. 2002, 102, 579-601.<br />
[2] B.G. Davis, Science 2004, 303, 480-482<br />
[3] S.I. van Kasteren, H.B. Kramer, H.H. Jensen, S.J. Campbell, J. Kirkpatrick,<br />
N.J. Oldham, D.C. Anthony, B.G. Davis, Nature 2007, 446, 1105-1109.<br />
[4] C. Fleming, A. Maldjian, D. Da Costa, P. Penny, R.C. Noble, N.R. Cameron,<br />
B.G. Davis, Nat. Chem. Biol. 2005, 1, 270-274<br />
[5] S.I. van Kasteren, H.B. Kramer, D.P. Gamblin, B.G. Davis, Nat. Protoc. 2007,<br />
2, 3185-3194<br />
[6] S.Y. Hong, G. Tobias, B. Ballesteros, F. El Oualid, J.C. Errey, K.J. Doores,<br />
A.I. Kirkland, P.D. Nellist, M.L.H. Green , B.G. Davis, J. Am. Chem. Soc. 2007,<br />
129, 10966-10967<br />
[7] G.J.L. Bernardes, J.M. Chalker, J.C. Errey, B.G. Davis, J. Am. Chem. Soc.<br />
2008, 130, 5052-5053
[8] S.I. van Kasteren, S.J. Campbell, S. Serres, D.C. Anthony, N.R. Sibson, B.G.<br />
Davis Proc. Natl Acad. Sci. U.S.A. 2009, 106, 18-23<br />
CARB 40<br />
Sialic acid recognition: Particles, chips, and cells<br />
Robert A. Field (1) , rob.field@bbsrc.ac.uk, Norwich Research Park, Colney Lane,<br />
Norwich Norfolk NR4 7UH, United Kingdom . (1) Department <strong>of</strong> Biological<br />
<strong>Chemistry</strong>, John Innes Centre, Norwich NR4 7UH, United Kingdom<br />
The sialic acids are a family <strong>of</strong> carbohydrates that are intimately associated with<br />
cellular recognition events. Using a combination <strong>of</strong> glycoarray and<br />
glyconanoparticle approaches, we have been investigating sialic acid recognition<br />
in the context <strong>of</strong> the human immune system and infection by the parasitic<br />
protozoan Trypanosoma cruzi. On one hand, glycoarray approaches with<br />
soluble, recombinant proteins have allowed us to explore similarities and<br />
differences in the carbohydrate-binding specificity <strong>of</strong> human and murine siglecs.<br />
On another, glyconanoparticle studies have enabled us ask questions about the<br />
structure and sialylation <strong>of</strong> parasite mucins. The combination <strong>of</strong> this information<br />
is now enabling us to begin to explore host recognition <strong>of</strong> parasite glycans<br />
through cell-based assays.<br />
CARB 41<br />
Functionalized nanoparticle for protein detection<br />
Chun-Cheng Lin (1) , cclin66@mx.nthu.edu.tw, 101, Sec. 2, Kuang Fu Rd.,<br />
Hsinchu Taiwan 30013, Taiwan Republic <strong>of</strong> China . (1) Department <strong>of</strong> <strong>Chemistry</strong>,<br />
National Tsing Hua University, Hsinchu Taiwan 30013, Taiwan Republic <strong>of</strong> China<br />
Biomolecule-conjugated nanoparticles (NPs) have been demonstrated to have<br />
promising applications on bioanalysis. Due to their large surface area to volume<br />
ratio and homogeneity in aqueous solution, functionalized nanoparticles were<br />
demonstrated as good carriers for affinity probes in target molecule separation.<br />
Our results showed that the ligand affinities with target proteins were significantly<br />
enhanced when ligands were assembled on the nanoparticles. The MALDI-TOF<br />
MS analysis was applied to identify the target molecules captured on the<br />
functionalized nanoparticles. In an attempt to fabricate highly active<br />
immunoprobes for serum biomarker detection, we developed a simple and<br />
effective method for site-specific and self-oriented immobilization <strong>of</strong> antibodies on<br />
magnetic nanoparticles (MNPs). The functionalized antibody conjugated<br />
nanoparticles were further extended as a multiplex probe to identify the<br />
biomarkers in human serum. The immobilization strategy was also applied to<br />
fabricate Fc-fused lectin microarray.
CARB 42<br />
Nanotechnology-based tools for the glycomics revolution: Silicon<br />
nanophotonic carbohydrate biosensors<br />
Daniel M Ratner (1) , dratner@u.washington.edu, Box 355061, Dept. <strong>of</strong><br />
Bioengineering, Seattle WA 98195, United States . (1) Department <strong>of</strong><br />
Bioengineering, University <strong>of</strong> Washington, Seattle WA 98195, United States<br />
Spurred by the post-genomic revolution in biological research, glycomics – the<br />
comprehensive study <strong>of</strong> carbohydrates (glycans) in biology – has undergone a<br />
rapid expansion, as researchers seek to unravel the many roles played by<br />
glycans in nature. This talk describes that application <strong>of</strong> nanotechnology and<br />
biointerfacial engineering to design new tools for glycomics research. Techniques<br />
including surface modification via molecular self-assembly, scanning probe<br />
microscopy characterization <strong>of</strong> glycosylated materials and an ultra-sensitive<br />
biosensing platform based on silicon nanophotonics will be discussed in the<br />
context <strong>of</strong> enabling glycomics research through nanoscale engineering.<br />
CARB 43<br />
Targeted glyco-magnetic nanoprobes for detection and molecular imaging<br />
<strong>of</strong> atherosclerosis<br />
Kheireddine El-Boubbou (1) , khb44@hotmail.com, Michigan State University,<br />
Department <strong>of</strong> <strong>Chemistry</strong>, East Lansing MI 48824, United States ; Medha<br />
Kamat (1) ; David Zhu (2) ; Ruiping Huang (2) ; George Abela (2) ; Xuefei Huang (1) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong>, Michigan State University, East Lansing MI 48823,<br />
United States (2) Department <strong>of</strong> Radiology, Michigan State University, East<br />
Lansing MI 48824, United States<br />
Cardiovascular diseases, <strong>of</strong>ten associated with atherosclerosis, are the leading<br />
cause <strong>of</strong> morbidity and mortality in the world. Despite the significant progress in<br />
cardiology, there remain large unmet needs to early detect atherosclerotic<br />
plaques, especially<br />
those which are prone to ruptures causing heart attacks and strokes. One <strong>of</strong> the<br />
major causes <strong>of</strong> such dramatic event is “inflammation” which occurs during early<br />
onset <strong>of</strong> the disease leading to over-expression <strong>of</strong> cell-adhesion glycoproteins.<br />
Our proposed work is based on the surveillance that hyaluronic acid (HA) is<br />
upregulated in atherosclerotic lesions and its principal cell-adhesion receptor,<br />
CD44 is involved in several atherogenic processes. Thus, we engineered novel<br />
hyaluronic superparamagnetic iron oxide nanoparticles (HA-SPIONPs) to be<br />
used not only as novel contrast cargo but also as pr<strong>of</strong>icient probes to noninvasively<br />
target atherosclerotic plaques using magnetic resonance imaging<br />
(MRI). The targeting agents on the external surface <strong>of</strong> the NPs will allow the<br />
selective labeling <strong>of</strong> the plaques. Due to the superparamagnetic nature <strong>of</strong> the
NPs, imaging <strong>of</strong> atherosclerotic plaques in rabbits was successfully examined.<br />
We anticipate that such novel nanoprobes will not only deepen our fundamental<br />
understanding <strong>of</strong> the molecular and cellular events characterizing unstable<br />
atherosclerotic plaques, but also be potentially developed into a highly innovative<br />
therapy for atherosclerosis.<br />
CARB 44<br />
<strong>Carbohydrate</strong>s on nanoparticles, surfaces and polymers: From basics to<br />
diagnostics applications<br />
Peter H. Seeberger (1) , peter.seeberger@mpikg.mpg.de, Arnimallee 22, Berlin<br />
14195, Germany . (1) Dept. <strong>of</strong> Biomolecular Systems, Max-Planck Institute for<br />
Colloids and Interfaces, Berlin 14195, Germany<br />
Cell surface oligosaccharides and glycosaminoglycans are important in signal<br />
transduction processes <strong>of</strong> biomedical significance. Described is an integrated<br />
platform relying on defined oligosacharides derived by automated solid phase<br />
syntehsis that are placed on many different nanoparticles, surfaces and<br />
polymers. By tuning the material as well as the carbohydrate, applications in the<br />
areas <strong>of</strong> diagnostics, vaccines and therapeutics. Particular focus will be placed<br />
on previously unpublished materials.<br />
For Previous work in our laboratory please see:<br />
1. Disney, M.D.; Zheng, J.; Swager, T.; Seeberger, P.H.; Visual Detection <strong>of</strong><br />
Bacteria with <strong>Carbohydrate</strong> Containing Fluorescent Polymers, J. Am. Chem. Soc.<br />
2004, 126, 13343-13346.<br />
2. de Paz, J.L.; Noti, C.; Boehm-Knoche, F.; Werner, S.; Seeberger, P.H.;<br />
Glycodendrimers Containing Synthetic Heparin Oligosaccharides to Regulate<br />
Heparin-Mediated Biological Processes; Chem. Biol. 2007, 14, 879-887.<br />
3. Kikkeri, R.; Lepenies, B.; Adibekian, A.;Laurino, P.; Seeberger, P.H.; In Vitro<br />
Imaging<br />
and In Vivo Liver Targeting with <strong>Carbohydrate</strong> Capped Quantum Dots; J. Am.
Chem. Soc. 2009, 131, 2110-2112.<br />
4. Kikkeri, R.; Gupta, T.; Hossain, L.H.; Kamena, F.; Gorodyska, G.; Beurer, E.;<br />
Textor, M.; Seeberger, P.H.; Ru(II)-Glycodendrimers as Probes to Study Lectin-<br />
<strong>Carbohydrate</strong> Interactions and Electrochemically Measure Mono- and<br />
Oligosaccharide Concentrations; Langmuir 2010, 26, 1520–1523.<br />
5. Kikkeri, R.; Laurino, P.; Odedra, A.; Seeberger, P.H.; Microreactor Synthesis<br />
<strong>of</strong> <strong>Carbohydrate</strong> Functionalized Quantum Dots; Angew. Chem. Int. Ed. 2010, 49,<br />
in press.<br />
CARB 45<br />
Efficient synthesis <strong>of</strong> siRNA-folic acid conjugates<br />
Rajendra Pandey (1) , rpandey@alnylam.com, 300 3rd St, 3rd Floor, Cambridge<br />
MA, United States ; Muthusamy Jayaraman (1) ; Anna Borodovsky (1) ; David<br />
Butler (1) ; Shigeo Matsuda (1) ; Gang Wang (1) ; Martin Maier (1) ; Bo Peng (1) ; Marjorie<br />
Solomon (1) ; Sergey Shulga-Morskoy (1) ; Klaus Charrise (1) ; David Bumcrot (1) ;<br />
Kristina Yucius (1) ; Victor Kotelianski (1) ; G. Rajeev Kallanthottathil (1) ; Muthiah<br />
Manoharan (1) . (1) Alnylam Pharmaceuticals, United States<br />
Cells and<br />
tissues have specific transport systems for nutrients like vitamins. Covalent<br />
conjugation <strong>of</strong> vitamin ligands will facilitate the delivery <strong>of</strong> siRNAs into<br />
cells having corresponding receptors and will enhance silencing <strong>of</strong> endogenous<br />
genes in vivo. Folate receptor/folic<br />
acid is a well-studied system with an affinity <strong>of</strong> 10 -10 M. The<br />
folate receptor is over-expressed in many types <strong>of</strong> tumor cells and in activated<br />
macrophages—cell types <strong>of</strong> relevance for disease treatment. Folic acid is an<br />
attractive choice for conjugation due to lack <strong>of</strong> immunogenicity and unlimited<br />
availability. Although folic acid has been employed as a targeting moiety for<br />
small molecule drugs and diagnostic agents, efficient synthetic methods to<br />
conjugate folic acid analogs to siRNAs were lacking. Part <strong>of</strong> the challenge<br />
arose due to the multiple functional groups in folic acid and incompatibility<br />
<strong>of</strong> deprotection conditions after automated RNA synthesis. We have prepared<br />
suitably protected folic acid phosphoramidites and solid supports with a<br />
variety <strong>of</strong> linkers. Using these building blocks, significant quantities <strong>of</strong><br />
folic acid conjugated siRNAs were synthesized. Synthesis, characterization, in<br />
vivo stability and uptake properties<br />
<strong>of</strong> these conjugates will be presented. <br />
CARB 46<br />
Combining 2'-TBDMS and 2'-ALE chemistry for on-column site specific<br />
modifications <strong>of</strong> RNA
Jeremy G Lackey (1) , jeremy.lackey@mcgill.ca, 801 Sherbrooke St. West,<br />
Montreal Quebec H3A 2K6, Canada ; Masad J Damha (1) ; Richard Johnsson (1) .<br />
(1) <strong>Chemistry</strong>, McGill University, Montreal Quebec H3A 2K6, Canada<br />
The celllular delivery and stability <strong>of</strong> siRNA must be overcome before it's<br />
therapeutic potential can be realized. Modifications at the 2'-position have been<br />
used extensively in this regard. Traditionally, 2'-modified nucleoside<br />
phosphoramidites are synthesized via a multi-step synthetic route and then<br />
incorporated in the oligonucleotide chain. This process can be tedious and<br />
expensive. Here in, we will describe a method for on-column site specific 2'modification.<br />
Various 2'-moieties will be introduced through the combination <strong>of</strong> 2'-<br />
ALE and 2'-TBDMS protecting groups. The characterization and biological<br />
activity <strong>of</strong> these molecules will also be discussed.<br />
CARB 47<br />
Synthesis and evaluation <strong>of</strong> bicyclic ketal-based cationic<br />
lipids for the delivery <strong>of</strong> siRNA via lipid nanoparticle delivery systems<br />
Muthusamy Jayaraman (1) , mJayaraman@alnylam.com, 300 3rd St, 3rd Floor,<br />
Cambridge MA, United States ; David Butler (1) ; Laxman Eltepu (1) ; Martin Maier (1) ;<br />
Tom Madden (2) ; Michael Hope (2) ; Ying Tam (2) ; Barbera Mui (2) ; Akin Akinc (1) ; Soma<br />
De (1) ; G. Rajeev Kallanthottathil (1) ; Muthiah Manoharan (1) ; Andrew Sprague (1) . (1)<br />
Alnylam Pharmaceuticals, United States (2) AlCana Technologies, Canada<br />
The<br />
discovery <strong>of</strong> RNA interference (RNAi) as a means to silence expression <strong>of</strong><br />
specific genes is potentially a great boon to modern medicine. Synthetic small<br />
interfering RNAs (siRNAs) are powerful agents for specifically eliciting the<br />
cleavage <strong>of</strong> mRNAs. A critical hurdle in realizing therapeutic application <strong>of</strong><br />
siRNA is it's cytosolic delivery across cellular membranes in vivo. Additionally,<br />
when administered systemically in<br />
unprotected form, nucleic acids suffer from poor pharmacokinetics due to rapid<br />
degradation by serum nucleases. Of methods tested so far, liposomal formulation<br />
has proven most successful in vivo. 1,
2,3 When administered intravenously in the form <strong>of</strong> an encapsulated<br />
lipid nanoparticle (LNP), siRNAs are delivered to the liver resulting in<br />
specific knockdown <strong>of</strong> hepatocytic targets.<br />
To identify the optimal LNPs for the delivery <strong>of</strong> siRNAs, cationic lipids<br />
derived from endo- and exo- cyclic aminodiols were evaluated. In this<br />
presentation we discuss design, synthesis and evaluation <strong>of</strong> the structure<br />
activity relationship <strong>of</strong> these cationic lipids in systemic delivery <strong>of</strong> siRNA.<br />
CARB 48<br />
Development <strong>of</strong> a stability-indicating, ion-pair RP-HPLC<br />
method for separation and quantitative determination <strong>of</strong> two siRNA<br />
duplexes in a<br />
liposome<br />
Veeravagu Murugaiah (1) , mmurugaiah@alnylam.com, 300 3rd St, 3rd Floor,<br />
Cambridge MA, United States ; William Zedalis (1) ; Gary Lavine (1) ; Klaus<br />
Charrise (1) ; Muthiah Manoharan (1) . (1) Alnylam Pharmaceuticals, United States<br />
We have developed a stability-indicating, ion-pair<br />
RP-HPLC method to separate and accurately quantitate two siRNA duplexes in a<br />
liposome without sample pretreatment.<br />
The first phase <strong>of</strong> the mobile phase system consisted <strong>of</strong> 385 mM<br />
hexafluoro-2-propanol, 14.5 mM triethylamine, and 5% methanol; the second<br />
consisted <strong>of</strong> 385 mM hexafluoro-2-propanol, 14.5 mM triethylamine, and 90%<br />
methanol. The column used was an XBridge<br />
C18 column (50 mm x 2.1 mm i.d., 2.5 µm particle size) and separation was<br />
performed at 60°C. Quantitation was<br />
achieved with UV detection at 260 nm. Linearity was established for the single<br />
strands <strong>of</strong> both siRNA duplexes for concentrations ranging from 10 to 110 µg/mL.<br />
Accuracy <strong>of</strong> the method was determined by<br />
replicate analysis (n=5) at four concentrations (R2 > 0.996 and RSDs <strong>of</strong> 1 –<br />
4%). Use <strong>of</strong> an ion pairing reagent<br />
compatible with mass spectrometric detection makes this method amenable to<br />
LC-MS impurity pr<strong>of</strong>iling.<br />
CARB 49<br />
Novel applications <strong>of</strong> aerosol-based detectors for the<br />
analysis <strong>of</strong> non-chromophore, multi-lipid, drug delivery vehicles<br />
William Zedalis (1) , bzedalis@alnylam.com, 300 3rd St, 3rd Floor, Cambridge<br />
MA, United States ; Matthias Kretschmer (1) ; Muthiah Manoharan (1) . (1) Alnylam<br />
Pharmaceuticals, United States
Analytical quantification <strong>of</strong> Lipid Nanoparticles (LNPs), a<br />
drug delivery vehicle consisting <strong>of</strong> multiple lipids, is challenging due to the<br />
lack <strong>of</strong> UV active chromophores on the constituents. A comparison employing<br />
three forms <strong>of</strong><br />
aerosol-based detection for liquid chromatography was performed using<br />
evaporative light-scattering detection (ELSD), charged aerosol detection (CAD),<br />
or condensation nucleation light-scattering detection (CNLSD). A summary <strong>of</strong> our<br />
findings will be presented.<br />
CARB 50<br />
Optimizing the LAL<br />
assay for detection <strong>of</strong> bacterial endotoxin in conjugated and formulated<br />
siRNAs<br />
Mara Broberg (1) , mbroberg@alnylam.com, 300 3rd St., 3rd Floor, Cambridge<br />
MA, United States ; Kathy Mills (1) ; Klaus Charisse (1) ; William Zedalis (1) ; Muthiah<br />
Manoharan (1) . (1) Alnylam Pharmaceuticals, United States<br />
Endotoxins, part <strong>of</strong> the outer cell wall <strong>of</strong> Gram negative<br />
bacteria, potently stimulate the innate immune response. The limulus<br />
amoebocyte<br />
lysate (LAL) test is the most widely used assay for detection <strong>of</strong> endotoxin and<br />
is an FDA requirement for testing <strong>of</strong> parental drug product safety. We used a<br />
commercially available LAL assay to detect and quantitatively determine the<br />
concentration <strong>of</strong> endotoxin in various conjugated siRNAs and liposomeformulated<br />
siRNAs. However, the lipids present in these various conjugates and<br />
formulations presented some challenges for the accurate determination <strong>of</strong><br />
endotoxin levels. We developed some solutions to overcome these problems and<br />
summary <strong>of</strong> our analytical processes will be presented.<br />
CARB 51<br />
Conjugation<br />
strategies for RNAs using copper-catalyzed click chemistry<br />
Chang Geng Peng (1) , ppeng@alnylam.com, 300 3rd St, 3rd Floor, Cambridge<br />
MA, United States ; Takeshi Yamada (1) ; Shigeo Matsuda (1) ; Haripriya Addepalli (1) ;<br />
Rowshon Alam (1) ; Narayanannair Jayaprakash (1) ; Muthusamy Jayaraman (1) ;<br />
David Butler (1) ; Rajendra Pandey (1) ; Kathy Mills (1) ; Martin Maier (1) ; Klaus<br />
Charrise (1) ; G. Rajeev Kallanthottathil (1) ; Muthiah Manoharan (1) . (1) Alnylam<br />
Pharmaceuticals, United States<br />
To<br />
improve the cellular uptake <strong>of</strong> siRNAs, we conjugated lipophilic molecules,
carbohydrates, and polyamines to appropriate sites <strong>of</strong> RNA molecules. A simple<br />
approach to achieve conjugation is via “click” chemistry involving the<br />
copper-catalyzed azide-alkyne cycloaddition reaction (CuAAC). We have<br />
achieved<br />
efficient synthesis <strong>of</strong> various conjugates by incorporating alkynes at different<br />
sites <strong>of</strong> the sugar residue <strong>of</strong> the ribonucleoside and reacting them with the<br />
desired ligands carrying an azido group. A summary <strong>of</strong> these synthetic processes<br />
are presented.<br />
CARB 52<br />
Non-nucleoside building blocks for copper-assisted<br />
and copper-free click chemistry for synthesis <strong>of</strong> oligonucleotide<br />
conjugates<br />
K.N. Jayaprakash (1) , jnair@alnylam.com, 300 3rd St, 3rd Floor, Cambridge MA,<br />
United States ; Chang G. Peng (1) ; Takeshi Yamada (1) ; David Butler (1) ; G Rajeev<br />
Kallanthottathil (1) ; Martin Maier (1) ; Muthiah Manoharan (1) . (1) Alnylam<br />
Pharmaceuticals, United States<br />
Cu(I)-mediated<br />
Huisgen 1,3-dipolar [3 + 2] cycloadditions between azides and alkynes,<br />
popularly known as “click reactions,” have been used for the conjugation <strong>of</strong><br />
number <strong>of</strong> biomolecules. We developed several non-nucleoside alkyne building<br />
blocks suitable for click chemistry based on hydroxyprolinol (see below) One <strong>of</strong><br />
the main issues in using copper mediated click chemistry for oligonucleotides<br />
and other bio-molecular conjugation is that trace amounts <strong>of</strong> potentially<br />
cytotoxic copper present in the product must be removed if these molecules are<br />
intended for therapeutic applications. Here, we report the use <strong>of</strong> copper-free<br />
click chemistry for the synthesis <strong>of</strong> oligonucleotide conjugates. Incorporation<br />
<strong>of</strong> various ligands into oligonucleotides in solution and solid phase will be<br />
presented.<br />
CARB 53<br />
Solid-support immobilized, reusable Cu (I)<br />
catalyst for "click reactions" <strong>of</strong> oligonucleotides with ligands<br />
Laxman Eltepu (1) , leltepu@alnylam.com, 300 3rd St, 3rd Floor, Cambridge MA,<br />
United States ; Chang G. Peng (1) ; K. Narayanannair Jayaprakash (1) ; Takeshi<br />
Yamada (1) ; Muthusamy Jayaraman (1) ; G. Rajeev Kallanthottathil (1) ; Muthiah<br />
Manoharan (1) . (1) Alnylam Pharmaceuticals, United States<br />
Cu(I)-catalyzed Huisgen's<br />
1,3-dipolar cycloaddition reaction, <strong>of</strong>ten referred as the “Click-reaction,” is<br />
<strong>of</strong>ten used in the bioconjugation <strong>of</strong> macromolecules due to its selectivity and
ease. One <strong>of</strong> the main limitations <strong>of</strong> this approach is that trace amounts <strong>of</strong><br />
cytotoxic Cu ions contaminate the products. Here, we report the synthesis <strong>of</strong> a<br />
Cu-catalyst on solid support and its application in “Click reactions” between<br />
small molecules and oligonucleotides. Use <strong>of</strong> this novel catalyst resulted in an<br />
experimentally simplified work-up protocol and prevented the leaching <strong>of</strong> the Cu<br />
metal. The catalyst could be re-cycled providing an environmentally clean<br />
process. The application <strong>of</strong> this novel Cu complex in “Click reactions” <strong>of</strong><br />
alkyne- and azide-bearing oligonucleotides or functionalized small molecules<br />
will be presented.<br />
CARB 54<br />
Synthesis <strong>of</strong> oligo spermine-containing<br />
oligonucleotides for siRNA delivery<br />
Shigeo Matsuda (1) , smatsuda@alnylam.com, 300 3rd St, 3rd Floor, Cambridge<br />
MA, United States ; Gang Wang (1) ; Ligang Zhang (1) ; Tianlei Lei (1) ; Rowshon<br />
Alam (1) ; Chang G Pang (1) ; K. N. Jayaprakash (1) ; Takeshi Yamada (1) ; David<br />
Butler (1) ; Maria Frank-Kamenetsky (1) ; Martin Maier (1) ; Klaus Charrise (1) ; Kevin<br />
Fitzgerald (1) ; G. Rajeev Kallanthottathil (1) ; Muthiah Manoharan (1) . (1) Alnylam<br />
Pharmaceuticals, United States<br />
To improve the cellular delivery <strong>of</strong><br />
siRNAs, we synthesized building blocks <strong>of</strong> multiunits <strong>of</strong> spermine derivatives<br />
and incorporated them into RNA oligonucleotides using phosphoramidite<br />
chemistry. Spermine-containing oligonucleotides were also synthesized by<br />
post-synthetic 'click' conjugation. A summary <strong>of</strong> these synthetic efforts for<br />
polyamine-conjugated siRNA synthesis are presented.<br />
CARB 55<br />
Allosteric modulation <strong>of</strong> DNA by minor groove binding polyamides<br />
David M Chenoweth (1) , dcheno@mit.edu, 77 Massachusetts Avenue, Bldg. 18-<br />
053, Cambridge MA 02139, United States ; Peter B Dervan (1) . (1) <strong>Chemistry</strong> and<br />
<strong>Chemical</strong> Engineering, California Institute <strong>of</strong> Technology, Pasadena CA 91125,<br />
United States<br />
Many human diseases are caused by dysregulated gene expression. The<br />
oversupply <strong>of</strong> transcription factors may be required for the growth and metastatic<br />
behavior <strong>of</strong> human cancers. Cell permeable small molecules that can be<br />
programmed to disrupt transcription factor-DNA interfaces could silence aberrant<br />
gene expression pathways. Pyrrole-imidazole polyamides are DNA minor groove<br />
binding molecules that are programmable for a large repertoire <strong>of</strong> DNA motifs.<br />
Atomic resolution X-ray crystal structures <strong>of</strong> Py/Im polyamides bound to DNA<br />
reveal a large widening <strong>of</strong> the minor groove and compression <strong>of</strong> the major groove
along with bending <strong>of</strong> the helix axis toward the major groove. These allosteric<br />
perturbations <strong>of</strong> the DNA helix provide a molecular basis for disruption <strong>of</strong><br />
transcription factor-DNA interfaces by small molecules, a minimum step in<br />
chemical control <strong>of</strong> gene networks.<br />
CARB 56<br />
Strong and selective molecular recognition <strong>of</strong> the DNA minor groove:<br />
Compound and DNA chemistry and unusual conformational matching<br />
W. David Wilson (1) , wdw@gsu.edu, 50 Decatur St, NSC, Atlanta GA 30303,<br />
United States ; Rupesh Nanjunda (1) ; Arvind Kumar (1) ; Manoj Munde (1) ; Yang<br />
Liu (1) ; Rebecca Hunt (1) ; David W. Boykin (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, Georgia<br />
State University, Atlanta GA, United States<br />
A number <strong>of</strong> organic cations recognize different DNA base sequences through<br />
minor groove complexes and have biological activity against diverse cells from<br />
cancer to infectious disease organisms. Although most classical minor groove<br />
binding agents specifically interact with AT base pairs, a combination <strong>of</strong> directed<br />
synthesis with a thorough set <strong>of</strong> biophysical methods has allowed us to discover<br />
new types <strong>of</strong> cooperative binding modes that provide for GC base pair<br />
recognition. The GC recognition modes involve a surprising stacking <strong>of</strong> dications<br />
that create an unprecedented tetracationic minor groove complex. Although<br />
design <strong>of</strong> minor groove binding agents to this time has been based on a<br />
restricted set <strong>of</strong> curvature and functional group position rules, we now realize that<br />
there are a number <strong>of</strong> other ways to target the groove. An overview <strong>of</strong> these new<br />
types <strong>of</strong> DNA complexes will be presented and contrasted with the classical AT<br />
specific complexes. Supported by NIH<br />
CARB 57<br />
Toward DNA recognition by a Janus Wedge approach<br />
Larry W McLaughlin (1) , mclaughl@bc.edu, 2609 Beacon St, Chestnut Hill MA<br />
02467, United States ; Ayan Pal (1) ; Han Chen (1) ; Meena Meena (1) . (1) Department<br />
<strong>of</strong> <strong>Chemistry</strong>, Boston College, Chestnut Hill MA 02467, United States<br />
Janus was the Roman god <strong>of</strong> beginnings and endings who is typically pictured<br />
with two faces. DNA recognition by a Janus-Wedge format involves the "wedge<br />
residue" inserting itself between the nucleobases <strong>of</strong> the Watson-Crick target and<br />
forming a DNA triplex. The J-W residues in the Janus Wedge triplex are<br />
composed <strong>of</strong> two hydrogen-bonding faces such that after insertion into the DNA<br />
duplex, typically through the major groove, the J-W residues are hydrogenbonded<br />
to both W-C faces <strong>of</strong> the target base pair. The product three-stranded<br />
complex contains significantly more interstrand hydrogen bonds than does the<br />
target W-C DNA duplex, and should be favored thermodynamically. A key
component <strong>of</strong> the recognition motif is to design residues that can discriminate all<br />
four base pairs, particularly A-T from T-A and G-C from C-G. Toward this end<br />
asymmetric residues will be described, each designed for recognition <strong>of</strong> one<br />
base pair.<br />
CARB 58<br />
Recognition <strong>of</strong> DNA major groove<br />
Dev P Arya (1) , dparya@clemson.edu, 461 Hunter, Clemson SC, United States ;<br />
Liang Xue (1) ; Sunil Kumar (1) . (1) <strong>Chemistry</strong>, Clemson University, Clemson SC<br />
29634, United States<br />
Sequence-specific recognition <strong>of</strong> duplex DNA can by achieved by small<br />
molecules using polyamides that target the DNA minor groove. The major groove<br />
<strong>of</strong> DNA is much richer in information content and a number <strong>of</strong> DNA-protein<br />
interactions take place in the major groove.<br />
In this seminar, I will present our results from efforts in targeting the DNA major<br />
groove. Our efforts towards the recognition <strong>of</strong> A and B form DNA will be<br />
presented.<br />
CARB 59<br />
Identification and cleavage site analysis <strong>of</strong> DNA sequences bound strongly<br />
by bleomycin<br />
Sidney M Hecht (1) , sid.hecht@asu.edu, Biodesign Institute, Tempe AZ 85287,<br />
United States . (1) Center for BioEnergetics, Biodesign Institute, Arizona State<br />
University, Tempe AZ 85287, United States<br />
A hairpin DNA library containing a randomized region was used to identify DNAs<br />
that bound strongly to bleomycin. Several strongly bound hairpin DNAs were<br />
identified and sequenced, and then used to characterize bleomycin-DNA<br />
interaction. Sequence analysis indicated that the randomized regions did not<br />
contain disproportionate numbers <strong>of</strong> 5'-GT-3' or 5'-GC-3' sequences, i.e. the DNA<br />
sequences cleaved predominantly when excess bleomycin is employed with<br />
arbitrarily closen DNAs. Further, the cleavage <strong>of</strong> these DNAs by Fe-bleomycin<br />
also involved atypical sequences, and involved a much larger proportion <strong>of</strong> alkali<br />
labile lesions than is observed when using excess bleomycin and arbitrarily<br />
chosen DNAs. The implications <strong>of</strong> these observations will be discussed.<br />
CARB 60<br />
Synthesis and characterization <strong>of</strong> chain-end functionalizable glycopolymer<br />
and its oriented glyco-macroligand formation
Satya Nandana Narla (1) , satyanandana.narla@gmail.com, 2121 Euclid Ave,<br />
Cleveland Ohio 44115, United States ; Xue-Long Sun (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, Cleveland State University, Cleveland Ohio 44115, United States<br />
Cell surface carbohydrates act as receptors for a variety <strong>of</strong> protein ligands and<br />
thereby play a significant role in a wide range <strong>of</strong> biological processes. Therefore,<br />
carbohydrate binding interaction by protein has provided a starting point for the<br />
development <strong>of</strong> new therapies and a new protein and cell isolation and targeting<br />
concept in biological research and applications. We report a facile synthesis <strong>of</strong><br />
α,ω-orthogonally functionalizable glycopolymer and its oriented glycomacrolignand<br />
formation for rapid purification and identification <strong>of</strong> carbohydratebinding<br />
protein applications. Specifically, α-biotin, ω-O-cyanate chain end<br />
functionalized glycopolymer was synthesized via cyanoxyl-mediated free-radical<br />
polymerization in one-pot fashion. Oriented glyco-macroligand formation and its<br />
glyco-affinity capturing <strong>of</strong> proteins were demonstrated by combining α-biotin endterminated<br />
glycopolymer with streptavidin modified magnetic beads and ω-Ocaynate<br />
end-terminatedglycipolymer with amine modified silica gel, respectively.<br />
The one-pot synthetic approach is expected to open a versatile pathway towards<br />
heterotelechelic (bio) functionalized polymers with defined chain end groups. The<br />
α,ω-orthogonally functionalizable glycopolymer can be used for both oriented<br />
polymer-protein conjugation and polymer surface immobilization for protein drug<br />
development and biosensor applications.<br />
CARB 61<br />
Separation and characterization <strong>of</strong> the glycosaminoglycan components <strong>of</strong><br />
proteoglycans<br />
Mellisa Ly (1) , lym@rpi.edu, 100 Nyroy Dr. Apt. 5, Troy NY 12180, United States ;<br />
Tatiana N Laremore (2) ; Kemal Solakyildirim (1) ; Robert J. Linhardt (1)(3)(4)(2) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic<br />
Institute, Troy NY 12180, United States (2) Center for Biotechnology and<br />
Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States (3) Department <strong>of</strong> Biology, Rensselaer Polytechnic Institute, Troy<br />
NY 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong> and Biological Engineering,<br />
Rensselaer Polytechnic Institute, Troy NY 12180, United States<br />
Proteoglycans (PGs) are a diverse group <strong>of</strong> glycoconjugates constituted by<br />
various core proteins post-translationally modified with linear, anionic<br />
glycosaminoglycans (GAGs) that consist <strong>of</strong> repeating disaccharide building<br />
blocks. Characterization <strong>of</strong> the GAG component <strong>of</strong> PGs is important because<br />
GAGs <strong>of</strong>ten dominate the physical, chemical, and biological properties <strong>of</strong> PGs.<br />
The structure <strong>of</strong> the chondroitin sulfate GAG chains <strong>of</strong> bikunin and decorin is<br />
investigated. Intact bikunin and decorin GAGs in a narrow molecular weight<br />
range were separated by continuous-elution preparative PAGE are analyzed<br />
using Fourier transform mass spectrometry. A method to elucidate the varying
sulfated domains for GAG chains <strong>of</strong> bikunin and decorin are developed using a<br />
combination <strong>of</strong> continuous-elution preparative PAGE, mass spectrometry, and<br />
GAG degrading enzymes, such as hyaluronidase (Streptococcus dysgalactiae)<br />
and exolytic chondroitin lyase II (Arthrobacter aurescens). Initial results indicate<br />
that there are regions <strong>of</strong> highly sulfated domains and nonsulfated domains.<br />
CARB 62<br />
Synthesis <strong>of</strong> nucleotide activated L-sugars from polyols via bioconversion<br />
Ryan D Woodyer (1) , rwoodyer@zuchem.com, 801 W Main St, Peoria IL 61606,<br />
United States . (1) zuChem, Inc., Chicago IL 60612, United States<br />
Many biologically active natural products owe their bioactivity at least in part to<br />
glycosylation. However, no commercially viable biological system exists for sitespecific<br />
attachment <strong>of</strong> L-sugar or modified-sugar monomers. We are developing<br />
methods for the production <strong>of</strong> activated l-sugars (in addition to other modified<br />
sugars) from inexpensive polyols using a combination <strong>of</strong> whole cell bioconversion<br />
and purified biocatalysts. L-Sugars, namely L-ribose, L-xylose, L-fucose, Lgalactose,<br />
L-gulose, were prepared from their inexpensive polyol counterparts<br />
using a recombinant whole cell E. coli biocatalyst containing and engineered<br />
polyol-1-dehydrogenase. Converting these L-sugars into nucleotide activated<br />
sugars is achieved using thermostable engineered biocatalysts, sugar-1-kinase<br />
and nucleotidyltransferase. Greater than 75% conversion to the corresponding<br />
sugar-1-P was achieved for L-glucose, L-arabinose, L-xylose, L-rhamnose, Lmannose,<br />
and 6-Azido-D-galactose using an engineered sugar-1-kinase. Gram<br />
scale synthesis <strong>of</strong> L-sugar-1-phosphates has been demonstrated with a spacetime<br />
yield <strong>of</strong> 200 g/L*d. Some activated sugars have been prepared using a<br />
nucleotidyltransferase and engineering to improve this step is currently<br />
underway.<br />
CARB 63<br />
Glycomorphology <strong>of</strong> the pulmonary vasculature: Endothelial cell<br />
glycocalyx and endothelial barrier function<br />
Eugene Ci<strong>of</strong>fi (1) , eaci<strong>of</strong>fi@jaguar1.usouthal.edu, 307 University Blvd. N., College<br />
<strong>of</strong> Medicine; MSB 3370, Mobile AL 36688-0002, United States . (1) Department<br />
<strong>of</strong> Pharmacology, University <strong>of</strong> South Alabama, Mobile AL 36688-0002, United<br />
States<br />
The endothelium forms a semi-permeable barrier between constituents <strong>of</strong> the<br />
blood and underlying tissue, consisting <strong>of</strong> glycoproteins, glycolipids and<br />
proteoglycans which coat the cell surface. The carbohydrate network that<br />
contributes to the glycocalyx is very complex, and its role in endothelial barrier<br />
function is poorly understood.
We have probed the molecular identities <strong>of</strong> endothelial cell surface<br />
carbohydrates using fluorescently-tagged lectins in a systematic way, revealing<br />
the presence <strong>of</strong> α-2,3-Gal and α-2,6-Gal/GalNAc terminal sialic acid residues that<br />
differed dependent upon the cell type. Overall a dramatic loss <strong>of</strong> cell-cell and cellmatrix<br />
adhesion was noted by the formation <strong>of</strong> large inter-endothelial cell gaps<br />
subsequent to a dose-dependent treatment with neuraminidases. In addition,<br />
endothelial barrier integrity was also evaluated using ECIS experiments, which<br />
correlate intracellular resistance with barrier function. Overall these observations<br />
suggest that the glycocalyx <strong>of</strong> endothelial cells contain terminal sialic acid<br />
residues that play a significant role in endothelial barrier function.<br />
CARB 64<br />
Multidimensional glycan arrays for enhanced lectin and antibody pr<strong>of</strong>iling<br />
Yalong Zhang (1) , Zhangy7@mail.nih.gov, 376 Boyles Street, Frederick Md<br />
21702, United States ; Qian Li (1) ; Christopher Campbell (1) ; Jeffrey Gildersleeve (1) .<br />
(1) National Cancer Institute, Frederick MD 21702, United States<br />
<strong>Carbohydrate</strong>-protein interactions are important in many biological processes.<br />
Multivalency is critical in the formation <strong>of</strong> a high avidity multivalent complex<br />
between carbohydrate ligands and the protein. Since many combinations <strong>of</strong> the<br />
structure, density, spacing and orientation <strong>of</strong> carbohydrate ligands are to be<br />
considered, identifying multivalent inhibitors and probes has been proven<br />
challenging. Here, we develop a new format array via spacing neoglycoproteins<br />
apart with unconjugated BSA. The array is composed <strong>of</strong> 591 combinations <strong>of</strong><br />
glycan structure and presentation. One application <strong>of</strong> the array was identifying<br />
inhibitors for five lectins. A key advantage is that multivalent inhibitors can be<br />
identified independent <strong>of</strong> the structure information. The other application was<br />
focus on the effects <strong>of</strong> neoglycoprotein density on antibody binding. We tested<br />
monoclonal antibodies, serum antibodies and serum antibodies induced by a<br />
prostate cancer vaccine. It was found that subpopulation antibodies could be<br />
distinguished, which could not have been detected otherwise.<br />
CARB 65<br />
Peptide nucleic acids bind strongly and sequence selectively to double<br />
helical RNA<br />
Thomas T Zengeya (1) , tzengey1@binghamton.edu, 4400 Vestal Parkway East,<br />
Binghamton New York 13902-6000, United States ; Ming Li (1) ; Eriks Rozners (1) .<br />
(1) <strong>Chemistry</strong>, State University <strong>of</strong> New York at Binghamton, Binghamton New<br />
York 13902, United States
Non-coding RNAs play a central role in gene expression, making them an<br />
attractive target for molecular recognition. However, because <strong>of</strong> RNA's<br />
conformational flexibility and the non-selective electrostatic interactions<br />
dominated by negatively charged phosphate backbone, discovery and designing<br />
<strong>of</strong> molecules that bind selectively to biologically and therapeutically relevant<br />
RNAs has been slow. Currently, most drugs that target RNA use hydrophobic<br />
and electrostatic interactions to achieve shape selective RNA recognition. For<br />
instance, ribosomal RNA is a receptor for antibiotics. However, while many<br />
molecules exhit high binding affinity, their selectivity is typically very low. In this<br />
presentation, we report that Peptide Nucleic Acids (PNA) bind strongly and<br />
sequence selectively to RNA duplexes. Our data suggests that the binding mode<br />
is formation <strong>of</strong> a Hoogsteen triple helix. In contrast to DNA, RNA triple helices<br />
have not been extensively studied. Moreover, stable triple helix formation<br />
between PNA and RNA has not been previously reported. Our results suggest<br />
that relatively small PNA analogues may be designed to recognize short<br />
homopurine stretches that are common in non-coding RNAs. It is conceivable<br />
that optimization <strong>of</strong> PNA properties using chemical modifications may provide a<br />
novel way to interfere with the function <strong>of</strong> non-coding RNAs.<br />
CARB 66<br />
Post surface function method for preparation <strong>of</strong> liposomal glycoconjugates<br />
Hailong Zhang (1) , zhldragon@gmail.com, 2121 Euclid Avenue, SI 414,<br />
Cleveland OH 44115, United States ; Yong Ma (1) ; Xue-Long Sun (1) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong>, Cleveland State University, Cleveland OH 44115,<br />
United States<br />
Cell surface glycans have been attractive biomimetic targets for potential<br />
biomedical applications since they act as receptors for a variety <strong>of</strong> protein ligands<br />
and are involved in a wide range <strong>of</strong> biological processes. Liposome, a spherical<br />
closed self-assembled lipid, has been extensively studied as model <strong>of</strong> cell<br />
membrane and mostly as carrier for drug/gene delivery application. Liposome<br />
surface functionalization with carbohydrate has been a versatile membrane<br />
mimetic approach and facilitates enormous potential applications <strong>of</strong> liposomes<br />
such as for glyco-model<br />
system, targeted drug and gene delivery as well as multivalent inhibitors.<br />
Conventional methods such as using amide or thiol-maleimide coupling as well<br />
as by imine or hydrazone linkage are less efficient and harmful for integrity and<br />
thus limit<br />
the liposome's biological application. Herein, we developed an efficient and<br />
chemoselective liposome surface glyco-functionalization method based on<br />
Staudinger ligation, in which carbohydrate derivative carrying a spacer with azide<br />
is conjugated onto the surface <strong>of</strong> preformed liposomes carrying a terminal<br />
triphosphine in PBS buffer (pH 7.4) and at room temperature. Specifically, by
using lactose and heparin as the model <strong>of</strong> carbohydrates, the effect <strong>of</strong> reaction<br />
conditions on integrity and stability <strong>of</strong> liposome was investigated by dynamic light<br />
scattering and the leakage <strong>of</strong> entrapped 5,6-carboxyfluorescein, respectively.<br />
Furthermore, the density and accessibility <strong>of</strong> grafted carbohydrate residues on<br />
the liposome surface were evaluated. The high specificity and high yield as well<br />
as biocompatible reaction condition natures<br />
<strong>of</strong> the Staudinger ligation approach make it an attractive alternative to all current<br />
protocols for liposome surface functionalization.<br />
CARB 67<br />
Approach to study carbohydrate-carbohydrate interactions involved in<br />
myelin using glycolipids assay in microtiter plate<br />
Jingsha Zhao (1) , Jingsha_zhao@brown.edu, 324 Brook St., Providence RI<br />
02912, United States ; Amit Basu (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, Brown<br />
University, Providence RI 02912, United States<br />
Glycolipid assay in the microtiter plate has been utilized to mimic the<br />
carbohydrate-carbohydrate interaction involved in myelin. As a model study,<br />
carbohydrate-lectin interaction was investigated with fluorescent lectins first. To<br />
study the carbohydrate-carbohydrate interaction, fluorescent silica nanoparticles,<br />
which have been functionalized with carbohydrates using the copper promoted<br />
azide-alkyne cycloaddition, was employed. The interaction between glycolipid in<br />
the plate and the carbohydrate on the surface <strong>of</strong> the fluorescent silica<br />
nanoparticles was elucidated by fluorescence spectrometer.<br />
CARB 68<br />
WITHDRAWN<br />
CARB 69
Synthesis and characterization <strong>of</strong> protein glycopolymer conjugate<br />
Valentinas Gruzdys (1) , v.gruzdys@csuohio.edu, 2121 Euclid Avenue, SR 382,<br />
Cleveland Ohio 44115-2214, United States ; Xue-Long Sun (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, Cleveland State University, Cleveland Ohio 44115, United States<br />
Covalent attachment <strong>of</strong> synthetic macromolecules is an effective way to improve<br />
protein stability with reduced immunogenicity and extended plasma half-lives. In<br />
addition, secondary property can be introduced, such as for protein targeted<br />
delivery. Glycoengineering aimed adding carbohydrates to proteins to alter their<br />
pharmacokinetic properties such as increasing in vivo activity and prolonging the<br />
duration <strong>of</strong> action have been a promising approach for protein therapeutics. In<br />
this presentation, a facile synthesis <strong>of</strong> chain end-functionalized glycopolymer for<br />
well-defined protein glycopolymer conjugation is reported. Briefly, an O-cyanate<br />
chain end-functionalized glycopolymer presenting multiple copies <strong>of</strong> lactose<br />
epitope units was synthesized via cyanoxyl-mediated free-radical polymerization<br />
in one-pot fashion. Bovine serum albumin (BSA) was employed as aminecontaining<br />
model protein for conjugation reaction, and the resulting proteinglycopolymer<br />
conjugate was characterized by SDS-PAGE. The versatility <strong>of</strong> the<br />
synthetic strategy presented in this work was the oriented multivalent<br />
carbohydrate modification <strong>of</strong> protein in straightforward approach and in aqueous<br />
conditions.<br />
CARB 70<br />
Robust analytical development for oligonucleotide manufacture<br />
Ipsita Roymoulik (1) , ipsita.roymoulik@aveciabiotech.com, 155 Fortune<br />
Boulevard, Milford MA 01757, United States . (1) Analytical Development, Avecia<br />
Oligomedicines, Milford MA 01757, United States<br />
A general overview <strong>of</strong> the role <strong>of</strong> Analytical Development in Oligonucleotide<br />
manufacture will be provided. Analytical techniques utilized to control the quality<br />
<strong>of</strong> oligonucleotides through robust purity and assay methods during in-process<br />
and QC release testing will be emphasized through understanding <strong>of</strong> typical<br />
oligonucleotide physico-chemical properties.<br />
The different categories <strong>of</strong> specifications and approaches to setting those<br />
specifications for critical quality attributes using risk and science based analytics<br />
will also be discussed.<br />
To ensure a back-up supply oligonucleotide therapeutic, sponsors require API<br />
through Drug Product quality being assessed at multiple laboratories and on<br />
multiple instrument platforms. Some <strong>of</strong> the key challenges due to the resulting<br />
analytical variability encountered during method qualifications and subsequent
validations will also be discussed through traditional versus statistical<br />
approaches.<br />
CARB 71<br />
Glycomics studies on central nervous system<br />
Fuming Zhang (1) , zhangf2@rpi.edu, 110 8th St., Troy NY 12180, United States ;<br />
Zhenling Liu (1) ; Kemal Solakyildirim (1) ; Dennis Pu (1) ; Robert J. Linhardt (1) . (1)<br />
Department <strong>of</strong> <strong>Chemical</strong> and Biological Engineering, Center for Biotechnology<br />
and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States<br />
Glycosaminoglycans (GAG) are a family <strong>of</strong> highly sulfated, complex,<br />
polydisperse linear polysaccharides that display a variety <strong>of</strong> important biological<br />
roles. They are known to participate in central nervous system processes such<br />
as development, cell migration, and neurite outgrowth. GAGs from swine brains<br />
and spinal cords were isolated and purified with a procedure including defat,<br />
proteolysis, anion-exchange chromatography; and methanol precipitation. The<br />
isolated GAGs were subjected to carbazole assay to quantify the amount <strong>of</strong> GAG<br />
in each tissue and polyacrylamide gel electrophoresis (PAGE) analysis. The<br />
disaccharide composition <strong>of</strong> chondroitin sulfate and heparan sulfate from the<br />
GAGs were determined using liquid chromotography and mass spectrometry<br />
(LC-MS). 1 H NMR spectroscopy was also used to characterize their structure.<br />
Finally the GAGs were biotinylated and immobilized on BIAcore SA biochips. The<br />
interactions between the GAGs and proteins related to central nervous system<br />
(such as midkine, brain-derived neurotrophic factor (BDNF) and Slit) were<br />
investigated using surface plasmon resonance (SPR).<br />
CARB 72<br />
Computational studies on the interactions <strong>of</strong> mannose with DOPC and<br />
POPC phospholipids<br />
Parthasarathi R (1) , rampartha@lanl.gov, parthasbioc@gmail.com, T6, MS K710,<br />
Los Alamos NM 87545, United States ; Gnanakaran S (1) . (1) Theoretical Biology<br />
& Biophysics Group, Los Alamos National Laboratory, Los Alamos NM 87545,<br />
United States<br />
Many pathogen induced potential causative immune responses are determined<br />
by the interaction <strong>of</strong> a virulence factor containing carbohydrates with membranes,<br />
including Mycobacterium tuberculosis in which its cell-wall glycolipid<br />
lipoarabinomannan (LAM), appear to be the most potent non-peptidic molecule to<br />
modulate the host immune response. In this study we seek a basic<br />
understanding <strong>of</strong> the nature <strong>of</strong> interactions between mannose and membranes<br />
since the initial molecular recognition <strong>of</strong> mannose molecules capping the LAM at
the host membrane is a critical step in the origin <strong>of</strong> pathogenesis. Relationships<br />
between physicochemical characteristics <strong>of</strong> mannose and its interactions with<br />
DOPC and POPC lipid molecules are investigated using density functional theory<br />
with 6-311++(d,p) basis sets. <strong>Carbohydrate</strong>-lipid interactions are probed with<br />
respect to competing hydrogen bonds to water. Based on these studies, relative<br />
strengths <strong>of</strong> mannose-water, lipid-water and mannose-lipid interactions are<br />
identified. Finally, we show that non-covalent interactions beyond hydrogen<br />
bonding may influence the stabilization <strong>of</strong> mannose at the membrane-water<br />
interface.<br />
CARB 73<br />
Optimizatin <strong>of</strong> analysis <strong>of</strong> glycosaminoglycans in biological samples<br />
Boyangzi Li (1) , lib4@rpi.edu, 110 8th Street, Troy NY 12180, United States ;<br />
Robert J Linhardt (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology,<br />
Rensselaer Polytechnic Institute, Troy NY 12180, United States<br />
As an important component <strong>of</strong> biological polysaccharides, glycosaminoglycans<br />
(GAGs) function as molecular co-receptors in numerous biological activities, such<br />
as cell–cell interaction, cell adhesion and migration, cell signaling, cell growth<br />
and differentiation. They mainly locate on the external side <strong>of</strong> cell membranes<br />
and contribute to the composition <strong>of</strong> extracellular matrix. Highly charged<br />
heterogeneous structure as they possess the sequence and composition <strong>of</strong><br />
GAGs in different biological samples and the same biological samples under<br />
different conditions are always in the interests <strong>of</strong> researchers to understand<br />
glycobiology. Here we present our work in optimization <strong>of</strong> the composition <strong>of</strong><br />
GAGs from biological samples—from improving extraction method to using<br />
different techniques for characterization, including reversed-phase ion-pair<br />
chromatography (R-IPC) coupled with electrospray ion mass spectroscopy (ESI-<br />
MS), strong anion exchange chromatography (SAX), capillary electrophoresis<br />
(CE) and introducing isotopically labeled disaccharide standards as internal<br />
standards for R-IPC coupled with ESI-MS.<br />
CARB 74<br />
Conformational analysis <strong>of</strong> nucleosides and nucleotides: PSEUROT 2010<br />
Steven M. Graham (1) , grahams@stjohns.edu, 8000 Utopia Parkway, Queens NY<br />
11439, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong>, St. John[apos]s University,<br />
Queens NY 11439, United States<br />
The biological activity <strong>of</strong> nucleosides and nucleotides is intimately tied to their<br />
conformation equilibrium. Most nucleosides and nucleotides exhibit a two-state<br />
'north-south' equilibrium. In this formalism, 'north' describes a nucleos(t)ide<br />
where C2' is puckered 'down' and C3' is puckered 'up'; 'south' describes a
nucleos(t)ide where C2' is puckered 'up' and C3' is puckered 'down'. The<br />
PSEUROT program allows a user to input 1 H- 1 H coupling constants, obtained<br />
from NMR spectroscopy, and to obtain as output the ratio <strong>of</strong> north/south<br />
conformers that would have produced the observed coupling constants. The<br />
original PSEUROT program, to our knowledge, has not been updated in a<br />
systematic fashion in the 30 years subsequent to its first description. This talk will<br />
focus on the merits <strong>of</strong> the use <strong>of</strong> molecular mechanics (AMBER) methods versus<br />
ab initio methods to update the PSEUROT program.<br />
CARB 75<br />
Role <strong>of</strong> DNA topography in recognition by proteins and small molecules<br />
Thomas D. Tullius (1) , tullius@bu.edu, 590 Commonwealth Avenue, Boston MA<br />
02215, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong> and Program in<br />
Bioinformatics, Boston University, Boston MA 02215, United States<br />
In the age <strong>of</strong> genomics, DNA is most <strong>of</strong>ten depicted as a<br />
string <strong>of</strong> letters. While this is useful for representing the large amount <strong>of</strong><br />
information encoded in a genome, the underlying molecular nature <strong>of</strong> DNA is<br />
obscured. Readout <strong>of</strong> genetic information is based on protein binding to<br />
specific sites in genomic DNA, but proteins cannot "read" DNA letters<br />
– they discriminate between potential DNA binding sites via the principles <strong>of</strong><br />
molecular recognition. To introduce a structural dimension to genome analysis,<br />
we developed a database <strong>of</strong> DNA structural patterns, ORChID, based on<br />
hydroxyl<br />
radical cleavage <strong>of</strong> DNA. We used ORChID to produce a topographical map <strong>of</strong><br />
the<br />
variation in DNA structure throughout the human genome, at single-nucleotide<br />
resolution. I will present recent<br />
work in which we use ORChID to assess how DNA topography contributes to the<br />
binding <strong>of</strong> proteins and small molecules to DNA.<br />
CARB 76<br />
Selective modulation <strong>of</strong> DNA polymerase activity by fixed-conformation<br />
nucleoside analogs<br />
Martin Egli (1) , martin.egli@vanderbilt.edu, School <strong>of</strong> Medicine, 607 Light Hall,<br />
Nashville Tennessee 37232-0146, United States ; Robert L. E<strong>of</strong>f (1) ; Victor E.
Marquez (2) ; F. Peter Guengerich (1) . (1) Department <strong>of</strong> Biochemistry and Center in<br />
Molecular Toxicology, Vanderbilt University, Nashville Tennessee 37232, United<br />
States (2) Laboratory <strong>of</strong> Medicinal <strong>Chemistry</strong>, Center for Cancer Research, NCI-<br />
Frederick, NIH, Frederick Maryland 21702, United States<br />
The ability to selectively modulate the activity <strong>of</strong> individual DNA polymerases<br />
(pols) or specific classes <strong>of</strong> pols is an attractive strategy for achieving therapeutic<br />
benefits such as anti-microbial activity. We have investigated the potential by<br />
various pols to selectively incorporate conformationally constrained dATP<br />
analogues with bicyclo[3.1.0]hexane (methanocarba-) pseudosugar moieties that<br />
are locked into either the N- or S-orientation. All pols tested showed a preference<br />
for the N- over the S-conformer, but with variable levels <strong>of</strong> inhibition relative to<br />
native dATP. Surprisingly, human pol iota showed increased activity towards the<br />
N-oriented conformer compared with dATP. Most notably, human pol eta was<br />
capable <strong>of</strong> inserting both conformers and is quite pr<strong>of</strong>icient at extension from the<br />
S-oriented conformer, in contrast to<br />
other polymerases tested. Our results expand upon the idea that individual pols<br />
utilize distinct mechanisms to achieve the same end and further our ability to<br />
develop novel nucleoside analogues that target specific enzymes.<br />
This work was supported by US National Institutes <strong>of</strong> Health grants R01<br />
ES010375 (F.P.G.), F32 CA119776 (R.L.E.), P30 ES000267 (F.P.G., M.E.), and<br />
P01 ES05355 (M.E.).<br />
CARB 77<br />
Structural mechanisms underlying DNA replication and human DNA<br />
mismatch repair<br />
Lorena S. Beese (1) , lsb@biochem.duke.edu, Nanaline Duke Bld., Rm 134;<br />
Research Dr.; Box 3711, Durham NC 27710, United States ; Jillian Orans (1) ;<br />
Weina Wang (1) ; Elizabeth McSweeney (1) ; Paul L Modrich (1) ; Quincy Tseng (1) . (1)<br />
Department <strong>of</strong> BIochemistry, Duke University Medical Center, Durham NC<br />
27705, United States<br />
Accurate DNA replication and repair are fundamental requirements for genomic<br />
stability. DNA polymerases achieve remarkably high replication fidelity, <strong>of</strong>ten<br />
allowing only a single insertion error for every 10,000 incorporated and<br />
additionally discriminating nearly a 1000- fold in favor <strong>of</strong> the 2'deoxyribonucleotide<br />
triphosphate (dNTP) compared to ribonucleotide<br />
triphosphate (rNTP). High-resolution crystal structures <strong>of</strong> DNA polymerase -<br />
substrate complexes provide new insights into mechanisms <strong>of</strong> nucleotide<br />
discrimination. Polymerase complexes with mismatched DNA <strong>of</strong>fer insight into a<br />
mechanisms for spontaneous mutation. DNA mismatch repair (MMR) further<br />
enhances the fidelity <strong>of</strong> replication, and initiates an apoptotic response to certain<br />
classes <strong>of</strong> DNA damage. Crystal structures <strong>of</strong> human MMR proteins MSH2-
MSH6, MSH2-MSH3, and exonuclease I bound to heteroduplex DNA lesions<br />
provide further insight into MMR and DNA recognition.<br />
CARB 78<br />
Manipulating the electrostatic potential in the DNA grooves: Effect on<br />
thermodynamic stability, ion binding, hydration and structure<br />
Barry Gold (1) , goldbi@pitt.edu, 512 Salk Hall, Pittsburgh PA 15216, United<br />
States . (1) Department <strong>of</strong> Pharmaceutical Sciences, University <strong>of</strong> Pittsburgh,<br />
Pittsburgh PA 15216, United States<br />
Cations, which sequence-selectively associate with DNA in both the major and<br />
minor grooves, play a significant role in determining DNA conformation and<br />
stability. In the major groove, cations are associated with the N7/O 6 edge <strong>of</strong><br />
guanine, while in the minor groove they are found at A-T pairs. Modification <strong>of</strong><br />
these potential cation binding sites should result in the reorganization <strong>of</strong> salts and<br />
water within the grooves, which in turn would affect local conformation and<br />
stability. We report the biophysical characterization <strong>of</strong> DNA duplexes in which we<br />
altered the N-7 position <strong>of</strong> purines (major groove) and the N-3 position <strong>of</strong> adenine<br />
(minor groove). These modifications either specifically eliminate a natural DNA<br />
cation binding sites or introduce a stable cationic appendage to mimic a high<br />
occupancy cation binding site. Interpretation <strong>of</strong> how these major and minor<br />
modifications affect DNA structure and stability, and potentially protein<br />
recognition <strong>of</strong> DNA, will be discussed.<br />
CARB 79<br />
Sequence-dependent recognition <strong>of</strong> minor groove width<br />
Barry Honig (1) , bh6@columbia.edu, 1130 St. Nicholas Ave, New York New York<br />
10032, United States . (1) Center for Computational Biology and Bioinformatics<br />
and Department <strong>of</strong> Biochemistry and Molecular Biophysics, Columbia University<br />
and Howard Hughes Medical Institute, New York New York 10032, United States<br />
By comprehensively analyzing the three dimensional structures <strong>of</strong> protein-DNA<br />
complexes, we show that the binding <strong>of</strong> arginines to narrow minor grooves is a<br />
widely used mode for protein-DNA recognition that is distinct from previously<br />
defined readout mechanisms. Minor groove narrowing is <strong>of</strong>ten associated with<br />
the presence <strong>of</strong> A-tracts, AT-rich sequences that exclude the flexible TpA step.<br />
The biophysical basis <strong>of</strong> this recognition mechanism involves an enhancement <strong>of</strong><br />
the negative potential <strong>of</strong> DNA, which is strongly correlated with minor groove<br />
width. Strikingly, arginines are frequently located at local minima in width and<br />
potential. These findings suggest that the ability to detect local variations in DNA<br />
shape and electrostatic potential is a general mechanism that enables proteins to<br />
use information in the minor groove, which otherwise <strong>of</strong>fers few opportunities for
the formation <strong>of</strong> base-specific hydrogen bonds, to achieve DNA binding<br />
specificity.<br />
CARB 80<br />
Glycosylation <strong>of</strong> substituted 4H-1,2,4-triazole-3-thiol<br />
El Sayed H. El Ashry (1) , eelashry60@hotmail.com, Faculty <strong>of</strong> Science,<br />
Alexandria Alexandria 31213, Egypt . (1) <strong>Chemistry</strong> Department, Alexandria<br />
University, Alexandria 31213, Egypt<br />
Glycosylsulfanyl-heterocycles have attracted much attention because <strong>of</strong> their<br />
biological activity and in particular inhibition <strong>of</strong> the activity <strong>of</strong> enzymes. Reaction<br />
<strong>of</strong> 5-substituted-4H-1,2,4-triazole-3-thiol with glycosyl halides in presence <strong>of</strong><br />
triethylamine gave the S-glycosides whereas the respective S,N 4 -bis(glycosyl)<br />
derivatives were synthesized in the presence <strong>of</strong> potassium carbonate. The S,N 2 -<br />
bis(glycosyl) isomer could also be isolated in minor amounts in some cases. MWI<br />
led to higher yields in much less time than the conventional methods and no<br />
change in regioselectivity has been noticed.<br />
CARB 81<br />
Expression and characterization <strong>of</strong> enzymes for bioenzymatic synthesis <strong>of</strong><br />
heparin<br />
Priscilla Paul (1) , paulp@rpi.edu, 110 8th Street, Troy NY 12180, United States ;<br />
Wenjing Zhao (1) ; Robert Linhardt (1) ; Jonathan Dordick (1) ; Jian Liu (2) . (1)<br />
Rensselaer Polytechnic Institute, Troy NY 12180, United States (2) University <strong>of</strong><br />
North Carolina, United States<br />
The goal <strong>of</strong> this project is to bioenzymatically synthesize 1 kg <strong>of</strong> non-animal<br />
source heparin that is chemically and biologically equivalent to United States<br />
Pharmacopoeia standard heparin. In the biosynthesis, N-sulfated heparosan,<br />
produced by E. coli fermentation and subsequent N-deacetylation and sulfation,<br />
is modified by several O-sulfotransferases (OST) to produce heparin. 2-OST, 6-<br />
OST-1, and 3-OST-1 transfer sulfo groups from 3'phosphoadenosine-<br />
5'phosphosulfate (PAPS) to the positions 2-O- <strong>of</strong> uronic acid, 6-O and 3-O <strong>of</strong><br />
glucosamine respectively. The expression and characterization <strong>of</strong> these<br />
necessary enzymes are critical to this process. OST have been expressed in E.<br />
coli and radiometrically assayed for their activities including 2-OST, 3-OST-1, 6-<br />
OST-1, and 6-OST-3. Sulfotransferase activity will be detected using continuous<br />
spectrophotometric coupled enzyme assays based on the regeneration <strong>of</strong> PAPS<br />
from desulfated 3'phosphoadenosine-5'phosphate (PAP) by recombinant aryl<br />
sulfotransferases. P-nitrophenyl sulfate will be the sulfate donor and visible<br />
spectrophotometric indicators <strong>of</strong> enzyme turnover will be utilized.
CARB 81<br />
Expression and characterization <strong>of</strong> enzymes for bioenzymatic synthesis <strong>of</strong><br />
heparin<br />
Priscilla Paul (1) , paulp@rpi.edu, 110 8th Street, Troy NY 12180, United States ;<br />
Wenjing Zhao (1) ; Robert Linhardt (1) ; Jonathan Dordick (1) ; Jian Liu (2) . (1)<br />
Rensselaer Polytechnic Institute, Troy NY 12180, United States (2) University <strong>of</strong><br />
North Carolina, United States<br />
The goal <strong>of</strong> this project is to bioenzymatically synthesize 1 kg <strong>of</strong> non-animal<br />
source heparin that is chemically and biologically equivalent to United States<br />
Pharmacopoeia standard heparin. In the biosynthesis, N-sulfated heparosan,<br />
produced by E. coli fermentation and subsequent N-deacetylation and sulfation,<br />
is modified by several O-sulfotransferases (OST) to produce heparin. 2-OST, 6-<br />
OST-1, and 3-OST-1 transfer sulfo groups from 3'phosphoadenosine-<br />
5'phosphosulfate (PAPS) to the positions 2-O- <strong>of</strong> uronic acid, 6-O and 3-O <strong>of</strong><br />
glucosamine respectively. The expression and characterization <strong>of</strong> these<br />
necessary enzymes are critical to this process. OST have been expressed in E.<br />
coli and radiometrically assayed for their activities including 2-OST, 3-OST-1, 6-<br />
OST-1, and 6-OST-3. Sulfotransferase activity will be detected using continuous<br />
spectrophotometric coupled enzyme assays based on the regeneration <strong>of</strong> PAPS<br />
from desulfated 3'phosphoadenosine-5'phosphate (PAP) by recombinant aryl<br />
sulfotransferases. P-nitrophenyl sulfate will be the sulfate donor and visible<br />
spectrophotometric indicators <strong>of</strong> enzyme turnover will be utilized.<br />
CARB 82<br />
Catalytic conversion <strong>of</strong> biomass-derived carbohydrates to useful chemicals<br />
in one step<br />
Weiran Yang (1) , wuy104@psu.edu, <strong>Chemistry</strong> Building, #439, University Park<br />
PA 16802, United States ; Ayusman Sen (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, The<br />
Pennsylvania State University, State College PA 16802, United States<br />
With diminishing fossil fuel reserves, the production <strong>of</strong> liquid fuels and valueadded<br />
chemicals directly from biomass is <strong>of</strong> great current interest. Here, we<br />
show that biomass derived carbohydrates such as fructose, glucose, and even<br />
cellulose can be catalytically converted to 2,5-dimethyltetrahydr<strong>of</strong>uran, 2methyltetrahydr<strong>of</strong>uran,<br />
2-methyltetrahydropyran, 2-ethyltetrahydr<strong>of</strong>uran, and 2methylcyclopentanone<br />
in one step. The total yield <strong>of</strong> these chemicals is relatively<br />
high. These chemicals are commercially important as solvents or starting<br />
materials. Also, since these liquid chemicals have high energy density and low<br />
solubility in water, they can be used as liquid fuels or fuel additives. A<br />
multifunctional catalyst composed <strong>of</strong> a transition metal-based component and an<br />
acid was employed.
CARB 82<br />
Catalytic conversion <strong>of</strong> biomass-derived carbohydrates to useful chemicals<br />
in one step<br />
Weiran Yang (1) , wuy104@psu.edu, <strong>Chemistry</strong> Building, #439, University Park<br />
PA 16802, United States ; Ayusman Sen (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, The<br />
Pennsylvania State University, State College PA 16802, United States<br />
With diminishing fossil fuel reserves, the production <strong>of</strong> liquid fuels and valueadded<br />
chemicals directly from biomass is <strong>of</strong> great current interest. Here, we<br />
show that biomass derived carbohydrates such as fructose, glucose, and even<br />
cellulose can be catalytically converted to 2,5-dimethyltetrahydr<strong>of</strong>uran, 2methyltetrahydr<strong>of</strong>uran,<br />
2-methyltetrahydropyran, 2-ethyltetrahydr<strong>of</strong>uran, and 2methylcyclopentanone<br />
in one step. The total yield <strong>of</strong> these chemicals is relatively<br />
high. These chemicals are commercially important as solvents or starting<br />
materials. Also, since these liquid chemicals have high energy density and low<br />
solubility in water, they can be used as liquid fuels or fuel additives. A<br />
multifunctional catalyst composed <strong>of</strong> a transition metal-based component and an<br />
acid was employed.<br />
CARB 83<br />
Behaviour <strong>of</strong> polysaccharide aggregates in asymmetrical<br />
field-flow fractionation and size-exclusion chromatography<br />
Leena Pitkänen (1) , leena.m.pitkanen@helsinki.fi, P.O. Box 27, Helsinki, Finland ;<br />
Maija Tenkanen (1) ; Päivi Tuomainen (1) . (1) Department <strong>of</strong> Food and<br />
Environmental Sciences, University <strong>of</strong> Helsinki, Helsinki 00014, Finland<br />
Asymmetrical flow field-flow fractionation (AsFlFFF) and high-performance sizeexclusion<br />
chromatography (HPSEC) are techniques for the separation and<br />
characterization <strong>of</strong> macromolecules; the latter more utilized so far for the analysis<br />
<strong>of</strong> hemicelluloses, such as wood- and cereal-derived xylans.<br />
In this paper, we compare the behavior <strong>of</strong> naturally occurring xylan aggregates in<br />
AsFlFFF and HPSEC and their effect on the obtained molar mass distribution<br />
and molecular characteristics (molar mass averages, size, intrinsic viscosity). We<br />
found out that aqueous xylan solutions contain two types <strong>of</strong> dense<br />
aggregates/molecules: assemblies with size above the size determined for<br />
individual xylan chains and some material having very low particle size.<br />
Although the amount <strong>of</strong> aggregates present in xylan solutions is extremely low,<br />
their role needs to be understood to avoid erroneous interpretation <strong>of</strong> AsFlFFF<br />
and HPSEC data. Using the data from both separation and detection systems
complementary information on the solution properties <strong>of</strong> aqueous polysaccharide<br />
solutions could be obtained.<br />
CARB 83<br />
Behaviour <strong>of</strong> polysaccharide aggregates in asymmetrical<br />
field-flow fractionation and size-exclusion chromatography<br />
Leena Pitkänen (1) , leena.m.pitkanen@helsinki.fi, P.O. Box 27, Helsinki, Finland ;<br />
Maija Tenkanen (1) ; Päivi Tuomainen (1) . (1) Department <strong>of</strong> Food and<br />
Environmental Sciences, University <strong>of</strong> Helsinki, Helsinki 00014, Finland<br />
Asymmetrical flow field-flow fractionation (AsFlFFF) and high-performance sizeexclusion<br />
chromatography (HPSEC) are techniques for the separation and<br />
characterization <strong>of</strong> macromolecules; the latter more utilized so far for the analysis<br />
<strong>of</strong> hemicelluloses, such as wood- and cereal-derived xylans.<br />
In this paper, we compare the behavior <strong>of</strong> naturally occurring xylan aggregates in<br />
AsFlFFF and HPSEC and their effect on the obtained molar mass distribution<br />
and molecular characteristics (molar mass averages, size, intrinsic viscosity). We<br />
found out that aqueous xylan solutions contain two types <strong>of</strong> dense<br />
aggregates/molecules: assemblies with size above the size determined for<br />
individual xylan chains and some material having very low particle size.<br />
Although the amount <strong>of</strong> aggregates present in xylan solutions is extremely low,<br />
their role needs to be understood to avoid erroneous interpretation <strong>of</strong> AsFlFFF<br />
and HPSEC data. Using the data from both separation and detection systems<br />
complementary information on the solution properties <strong>of</strong> aqueous polysaccharide<br />
solutions could be obtained.<br />
CARB 84<br />
Key new observations in the synthesis <strong>of</strong> thiosialosides<br />
Robert A Falconer (1) , r.a.falconer1@bradford.ac.uk, Richmond Road, Bradford<br />
West Yorkshire BD7 1DP, United Kingdom ; Ines F Oliveira (1) ; Goreti R Morais (1) ;<br />
Bradley R Springett (1) . (1) Institute <strong>of</strong> Cancer Therapeutics, School <strong>of</strong> Life<br />
Sciences, University <strong>of</strong> Bradford, Bradford BD7 1DP, United Kingdom<br />
The disulfide bond plays an important role in both chemistry and biology, being<br />
responsible for the formation <strong>of</strong> higher order structures in peptides and proteins.<br />
Glycosyl disulfides are also useful tools to aid understanding <strong>of</strong> enzyme systems<br />
and are <strong>of</strong> interest as donors in glycosylation reactions.<br />
In our quest to build a library <strong>of</strong> thiosialosides via the simultaneous selective Sdeacetylation<br />
<strong>of</strong> 2-sialyl thioacetate and alkylation, disulfides were observed.
Further experiments demonstrated that such disulfides resulted from the<br />
alkylation <strong>of</strong> a sialyl acetyl disulfide, identified as a randomly constant by-product<br />
during the synthesis <strong>of</strong> thiosialosides. Formation <strong>of</strong> this disulfide by-product is<br />
extremely variable and dependent on the commercial source <strong>of</strong> KSAc.<br />
We describe our efforts to identify and characterise this by-product, and our<br />
efforts towards understanding the mechanism <strong>of</strong> its formation.<br />
CARB 85<br />
Synthesis and structural optimization <strong>of</strong> antifungal kanamycin B analogs<br />
Marina Y FOSSO (1) , marina.fosso@aggiemail.usu.edu, 0300 Old Main Hill, Box<br />
# 72, Logan UT 84322, United States ; Yukie KAWASAKI (2) ; Sanjib<br />
SHRESTHA (2) ; Jon TAKEMOTO (2) ; Tom C.-W. CHANG (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong> and Biochemistry, Utah State University, Logan UT 84322, United<br />
States (2) Department <strong>of</strong> Biology, Utah State University, Logan UT 84322, United<br />
States<br />
Besides their traditional role <strong>of</strong> antibacterial agents, aminoglycosides, both<br />
classical and structurally unusual ones, have been reported to exhibit antifungal<br />
activity. This was confirmed as several members <strong>of</strong> our library <strong>of</strong> kanamycin B<br />
analogs were found to possess significant antifungal activity, notably against<br />
Rhodotorula piliminae and Fusarium graminearum, the latter being the major<br />
causal agent <strong>of</strong> wheat head blight and maize ear rot in North America. Based on<br />
the leads and following the synthesis <strong>of</strong> two new kanamycin B analogs, we were<br />
able to draw a structure – activity relationship which reveals that attaching an 8carbon<br />
alkyl chain at position O-4´´ <strong>of</strong> kanamycin B converts it into a fungicide<br />
with loss <strong>of</strong> antibacterial activity. Further effort has then directed to the<br />
investigation <strong>of</strong> the mechanism <strong>of</strong> action and to the optimization <strong>of</strong> the synthetic<br />
protocol.<br />
CARB 85<br />
Synthesis and structural optimization <strong>of</strong> antifungal kanamycin B analogs<br />
Marina Y FOSSO (1) , marina.fosso@aggiemail.usu.edu, 0300 Old Main Hill, Box<br />
# 72, Logan UT 84322, United States ; Yukie KAWASAKI (2) ; Sanjib<br />
SHRESTHA (2) ; Jon TAKEMOTO (2) ; Tom C.-W. CHANG (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong> and Biochemistry, Utah State University, Logan UT 84322, United<br />
States (2) Department <strong>of</strong> Biology, Utah State University, Logan UT 84322, United<br />
States<br />
Besides their traditional role <strong>of</strong> antibacterial agents, aminoglycosides, both<br />
classical and structurally unusual ones, have been reported to exhibit antifungal<br />
activity. This was confirmed as several members <strong>of</strong> our library <strong>of</strong> kanamycin B
analogs were found to possess significant antifungal activity, notably against<br />
Rhodotorula piliminae and Fusarium graminearum, the latter being the major<br />
causal agent <strong>of</strong> wheat head blight and maize ear rot in North America. Based on<br />
the leads and following the synthesis <strong>of</strong> two new kanamycin B analogs, we were<br />
able to draw a structure – activity relationship which reveals that attaching an 8carbon<br />
alkyl chain at position O-4´´ <strong>of</strong> kanamycin B converts it into a fungicide<br />
with loss <strong>of</strong> antibacterial activity. Further effort has then directed to the<br />
investigation <strong>of</strong> the mechanism <strong>of</strong> action and to the optimization <strong>of</strong> the synthetic<br />
protocol.<br />
CARB 86<br />
E. coli K5 heparosan production for bioengineered heparin<br />
Zhenyu Wang (1)(2) , wangz2@rpi.edu, Room 4005, Troy NY 12180, United States<br />
; Mellisa Ly (3)(2) ; Fuming Zhang (4)(2) ; Zhenqing Zhang (5) ; Jonathan S.<br />
Dordick (1)(4)(2) ; Robert J. Linhardt (1)(4)(3)(2) . (1) Department <strong>of</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy NY 12180, United States (2) Center for Biotechnology<br />
and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States (3) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy NY 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong><br />
and Biological Engineering, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States (5) Baxter International Inc., United States<br />
Heparosan is an acidic polysaccharide natural product. It is a critical precursor in<br />
heparin biosynthesis and in the chemoenzymatic synthesis <strong>of</strong> bioengineered<br />
heparin. The process <strong>of</strong> heparosan production by E. coli K5 fermentation<br />
significantly affects the cost and the pharmacological properties <strong>of</strong> the final<br />
product bioengineered heparin. In this study, heparosan produced by E. coli K5<br />
fermentation was characterized and the fermentation process was optimized and<br />
controlled to produce the ideal bioengineered heparin precursor. E. coli strain<br />
improvement by metabolic engineering and gene manipulation was also<br />
investigated.<br />
CARB 86<br />
E. coli K5 heparosan production for bioengineered heparin<br />
Zhenyu Wang (1)(2) , wangz2@rpi.edu, Room 4005, Troy NY 12180, United States<br />
; Mellisa Ly (3)(2) ; Fuming Zhang (4)(2) ; Zhenqing Zhang (5) ; Jonathan S.<br />
Dordick (1)(4)(2) ; Robert J. Linhardt (1)(4)(3)(2) . (1) Department <strong>of</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy NY 12180, United States (2) Center for Biotechnology<br />
and Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States (3) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy NY 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong>
and Biological Engineering, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States (5) Baxter International Inc., United States<br />
Heparosan is an acidic polysaccharide natural product. It is a critical precursor in<br />
heparin biosynthesis and in the chemoenzymatic synthesis <strong>of</strong> bioengineered<br />
heparin. The process <strong>of</strong> heparosan production by E. coli K5 fermentation<br />
significantly affects the cost and the pharmacological properties <strong>of</strong> the final<br />
product bioengineered heparin. In this study, heparosan produced by E. coli K5<br />
fermentation was characterized and the fermentation process was optimized and<br />
controlled to produce the ideal bioengineered heparin precursor. E. coli strain<br />
improvement by metabolic engineering and gene manipulation was also<br />
investigated.<br />
CARB 87<br />
Stochastic simulation <strong>of</strong> lectin microarrays with nanosensor transducers:<br />
Potential platforms for optimal, high-throughput screening and pr<strong>of</strong>iling <strong>of</strong><br />
glycoproteins<br />
Nigel F Reuel (1) , reuel@mit.edu, 77 Massachusetts Avenue, Room 66-566,<br />
Cambridge MA 02139, United States ; Jin-Ho Ahn (1) ; Michael S. Strano (1) . (1)<br />
Department <strong>of</strong> <strong>Chemical</strong> Engineering, Massachusetts Institute <strong>of</strong> Technology,<br />
Cambridge MA 02139, United States<br />
As expressed, glycoprotein therapeutics expand in market share, there is a<br />
growing need for high-throughput platforms for pr<strong>of</strong>iling (to determine their<br />
homogeneity) and screening (to detect any deleterious glycan additions). An<br />
array <strong>of</strong> carefully selected lectins could provide such a platform. A Kinetic Monte<br />
Carlo method has been employed to model the interactions <strong>of</strong> glycoproteins with<br />
a lectin array to determine which lectins would be <strong>of</strong> best use for screening and<br />
pr<strong>of</strong>iling. Interaction parameters between many lectins and glycans were<br />
supplied as dissociation constants (KD). Clear contrast for screening purposes is<br />
manifested in both frequency response and average site occupancy. Three<br />
current applications <strong>of</strong> the array are modeled: 1) screening protein therapeutics,<br />
2) differentiating arthritic disease, and 3) evaluating cancer biomarkers. The<br />
complete pr<strong>of</strong>iling <strong>of</strong> glycoproteins without a priori knowledge <strong>of</strong> their synthesis is<br />
further discussed, as well as the application <strong>of</strong> stochastic nanosensors to label<br />
free detection.<br />
CARB 87<br />
Stochastic simulation <strong>of</strong> lectin microarrays with nanosensor transducers:<br />
Potential platforms for optimal, high-throughput screening and pr<strong>of</strong>iling <strong>of</strong><br />
glycoproteins
Nigel F Reuel (1) , reuel@mit.edu, 77 Massachusetts Avenue, Room 66-566,<br />
Cambridge MA 02139, United States ; Jin-Ho Ahn (1) ; Michael S. Strano (1) . (1)<br />
Department <strong>of</strong> <strong>Chemical</strong> Engineering, Massachusetts Institute <strong>of</strong> Technology,<br />
Cambridge MA 02139, United States<br />
As expressed, glycoprotein therapeutics expand in market share, there is a<br />
growing need for high-throughput platforms for pr<strong>of</strong>iling (to determine their<br />
homogeneity) and screening (to detect any deleterious glycan additions). An<br />
array <strong>of</strong> carefully selected lectins could provide such a platform. A Kinetic Monte<br />
Carlo method has been employed to model the interactions <strong>of</strong> glycoproteins with<br />
a lectin array to determine which lectins would be <strong>of</strong> best use for screening and<br />
pr<strong>of</strong>iling. Interaction parameters between many lectins and glycans were<br />
supplied as dissociation constants (KD). Clear contrast for screening purposes is<br />
manifested in both frequency response and average site occupancy. Three<br />
current applications <strong>of</strong> the array are modeled: 1) screening protein therapeutics,<br />
2) differentiating arthritic disease, and 3) evaluating cancer biomarkers. The<br />
complete pr<strong>of</strong>iling <strong>of</strong> glycoproteins without a priori knowledge <strong>of</strong> their synthesis is<br />
further discussed, as well as the application <strong>of</strong> stochastic nanosensors to label<br />
free detection.<br />
CARB 88<br />
Evaluation <strong>of</strong> different thioesters for glycocluster synthesis applying native<br />
chemical ligation<br />
Johannes W. Wehner (1) , jwehner@oc.uni-kiel.de, Otto-Hahn-Platz 4, Kiel<br />
Schleswig-Holstein 24098, Germany ; Thisbe K. Lindhorst (1) . (1) Otto Diels<br />
Institute <strong>of</strong> Organic <strong>Chemistry</strong>, Christiana Albertina University <strong>of</strong> Kiel, Kiel 24098,<br />
Germany<br />
Multivalent glycomimetics such as glycoclusters and glycodendrimers [1] are<br />
valuable tools for structure-function investigations in the glyco sciences.[2]<br />
During the last decade, our group has utilized various chemistries to achieve<br />
glycoclusters <strong>of</strong> different architectures and has tested their biology mainly in<br />
bacterial adhesion.[3] In this work it has been the aim to evaluate native chemical<br />
ligation (NCL) for the synthesis <strong>of</strong> clustered glycosides. This approach is<br />
appealing, because it does not require protecting groups and can be extended to<br />
more complex targets, eventually. Native chemical ligation was first reported in<br />
1953 [4] and utilized in 1994 by S. B. Kent et al. to synthesize peptides [5] and<br />
has then been developed into a popular chemoselective method for ligation <strong>of</strong><br />
large peptides and glycopeptide fragments.[6]<br />
Glycocluster synthesis via NCL requires a glycocysteine derivative, which can be<br />
reacted with a thioester in a subsequent ligation step. Thus, a number <strong>of</strong><br />
thioesters with varying polarities (type 1 - 4) was synthesized and ligated with a<br />
mannosidic cysteine derivative.
[1] N. Röckendorf; Th. K. Lindhorst, Top. Curr. Chem. 2001, 217, 201-238; [2] M.<br />
Lahmann, Top. Curr. Chem. 2009, 288, 17-65; [3] M. Dubber; O. Sperling; Th. K.<br />
Lindhorst, Org. Biomol. Chem. 2006, 4, 3913-3922; [4] T. Wieland; E.<br />
Bokelmann; L. Bauer; H. U. Lang; H. Lau, Liebigs Ann. 1953, 583, 129-149; [5]<br />
P. E. Dawson; T. W. Muir; I. Clark-Lewis; S. B. Kent, Science 1994, 266, 776-<br />
779; [6] C. P. R. Hackenberger; D. Schwarzer, Angew. Chem. 2008, 120, 10182-<br />
10228; Angew. Chem. Int. Ed. 2008, 47, 10030-10074.<br />
CARB 88<br />
Evaluation <strong>of</strong> different thioesters for glycocluster synthesis applying native<br />
chemical ligation<br />
Johannes W. Wehner (1) , jwehner@oc.uni-kiel.de, Otto-Hahn-Platz 4, Kiel<br />
Schleswig-Holstein 24098, Germany ; Thisbe K. Lindhorst (1) . (1) Otto Diels<br />
Institute <strong>of</strong> Organic <strong>Chemistry</strong>, Christiana Albertina University <strong>of</strong> Kiel, Kiel 24098,<br />
Germany<br />
Multivalent glycomimetics such as glycoclusters and glycodendrimers [1] are<br />
valuable tools for structure-function investigations in the glyco sciences.[2]<br />
During the last decade, our group has utilized various chemistries to achieve<br />
glycoclusters <strong>of</strong> different architectures and has tested their biology mainly in<br />
bacterial adhesion.[3] In this work it has been the aim to evaluate native chemical<br />
ligation (NCL) for the synthesis <strong>of</strong> clustered glycosides. This approach is<br />
appealing, because it does not require protecting groups and can be extended to<br />
more complex targets, eventually. Native chemical ligation was first reported in<br />
1953 [4] and utilized in 1994 by S. B. Kent et al. to synthesize peptides [5] and<br />
has then been developed into a popular chemoselective method for ligation <strong>of</strong><br />
large peptides and glycopeptide fragments.[6]<br />
Glycocluster synthesis via NCL requires a glycocysteine derivative, which can be<br />
reacted with a thioester in a subsequent ligation step. Thus, a number <strong>of</strong><br />
thioesters with varying polarities (type 1 - 4) was synthesized and ligated with a<br />
mannosidic cysteine derivative.
[1] N. Röckendorf; Th. K. Lindhorst, Top. Curr. Chem. 2001, 217, 201-238; [2] M.<br />
Lahmann, Top. Curr. Chem. 2009, 288, 17-65; [3] M. Dubber; O. Sperling; Th. K.<br />
Lindhorst, Org. Biomol. Chem. 2006, 4, 3913-3922; [4] T. Wieland; E.<br />
Bokelmann; L. Bauer; H. U. Lang; H. Lau, Liebigs Ann. 1953, 583, 129-149; [5]<br />
P. E. Dawson; T. W. Muir; I. Clark-Lewis; S. B. Kent, Science 1994, 266, 776-<br />
779; [6] C. P. R. Hackenberger; D. Schwarzer, Angew. Chem. 2008, 120, 10182-<br />
10228; Angew. Chem. Int. Ed. 2008, 47, 10030-10074.<br />
CARB 89<br />
NMR for structure elucidation <strong>of</strong> commercially available heparin<br />
polysaccharides<br />
Kemal Solakyildirim (1) , solakk@rpi.edu, 2100 Massachusetts Ave. Apt#2A, Troy<br />
New York 12180, United States ; Scott A. McCallum (2) ; Robert J. Linhardt (1)(2)(3)(4) .<br />
(1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic<br />
Institute, Troy New York 12180, United States (2) Center for Biotechnology and<br />
Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy New York 12180,<br />
United States (3) Department <strong>of</strong> Biology, Rensselaer Polytechnic Institute, Troy<br />
New York 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong> and Biological<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States<br />
Heparin is highly negatively charged, heterogeneous linear polysaccharide that is<br />
mainly used in pharmaceutical products as an anticoagulant drug for the<br />
treatment <strong>of</strong> thrombosis, thrombophlebitis, and embolism. Commercial USP<br />
heparin is primarily obtained from porcine intestinal mucosa. Recently, heparin<br />
was contaminated with oversulfated chondroitin sulfate (OSCS), which caused<br />
the death <strong>of</strong> nearly 100 <strong>American</strong>s. However, it may now possible to make a<br />
bioengineered heparin from non-animal sources. FDA approval would require a<br />
bioengineered heparin to have a structure identical to USP heparin. NMR<br />
spectroscopy is a powerful and high-resolution technique capable <strong>of</strong> detailed<br />
structural elucidation <strong>of</strong> the heparin polysaccharide. Commercial heparin<br />
samples, obtained from various manufacturers around the world, were analyzed<br />
using 1D 1 H and 13 C NMR and 2D H-H COSY, TOCSY, HMQC, NOESY NMR<br />
experiments. The specific aim <strong>of</strong> this study is to use NMR to establish the<br />
structural features present in USP heparin.<br />
CARB 89<br />
NMR for structure elucidation <strong>of</strong> commercially available heparin<br />
polysaccharides
Kemal Solakyildirim (1) , solakk@rpi.edu, 2100 Massachusetts Ave. Apt#2A, Troy<br />
New York 12180, United States ; Scott A. McCallum (2) ; Robert J. Linhardt (1)(2)(3)(4) .<br />
(1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic<br />
Institute, Troy New York 12180, United States (2) Center for Biotechnology and<br />
Interdisciplinary Studies, Rensselaer Polytechnic Institute, Troy New York 12180,<br />
United States (3) Department <strong>of</strong> Biology, Rensselaer Polytechnic Institute, Troy<br />
New York 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong> and Biological<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States<br />
Heparin is highly negatively charged, heterogeneous linear polysaccharide that is<br />
mainly used in pharmaceutical products as an anticoagulant drug for the<br />
treatment <strong>of</strong> thrombosis, thrombophlebitis, and embolism. Commercial USP<br />
heparin is primarily obtained from porcine intestinal mucosa. Recently, heparin<br />
was contaminated with oversulfated chondroitin sulfate (OSCS), which caused<br />
the death <strong>of</strong> nearly 100 <strong>American</strong>s. However, it may now possible to make a<br />
bioengineered heparin from non-animal sources. FDA approval would require a<br />
bioengineered heparin to have a structure identical to USP heparin. NMR<br />
spectroscopy is a powerful and high-resolution technique capable <strong>of</strong> detailed<br />
structural elucidation <strong>of</strong> the heparin polysaccharide. Commercial heparin<br />
samples, obtained from various manufacturers around the world, were analyzed<br />
using 1D 1 H and 13 C NMR and 2D H-H COSY, TOCSY, HMQC, NOESY NMR<br />
experiments. The specific aim <strong>of</strong> this study is to use NMR to establish the<br />
structural features present in USP heparin.<br />
CARB 90<br />
Syntheses <strong>of</strong> C-5-spirocyclic C-glycoside SGLT2 inhibitors<br />
Benjamin A. Thuma (1) , benjamin.thuma@pfizer.com, Eastern Point Rd., Groton<br />
CT 06340, United States ; Ralph P. Robinson (1) ; Cathy Préville (1) ; Matthew R.<br />
Reese (2) ; Robert J. Maguire (1) ; Christopher L. Carr (2) ; Michael T. Leininger (2) ;<br />
André Lowe (2) ; Claire M. Steppan (2) ; Vincent Mascitti (1) . (1) CVMED Medicinal<br />
<strong>Chemistry</strong>, Pfizer Global Research and Development, Groton CT 06340, United<br />
States (2) Pfizer Global Research and Development, Groton CT 06340, United<br />
States<br />
SGLT2 inhibitors theoretically should promote weight loss and control<br />
hyperglycemia in a glucose-dependent yet insulin-independent manner, thereby<br />
potentially making such therapeutic agents very compelling complements to the<br />
current arsenal <strong>of</strong> anti-diabetic agents. We recently reported that some C-5spirocyclic<br />
C-glycoside derivatives are potent and selective SGLT2 inhibitors. 1 In<br />
this poster, several syntheses <strong>of</strong> C-5-spirocycle-containing C-glycosides are<br />
presented. Particularly, a multigram-scale synthesis capitalizing on a one pot<br />
aldol-Cannizzaro sequence is described. Spiro oxetane formation using an<br />
unprotected penta-ol C-glycoside as substrate is also exemplified. 2
1. Robinson, R. P. et al. Bioorg. Med. Chem. Lett. 2010, 20, 1569.<br />
2. Mascitti, V. et al. Tetrahedron Letters 2010, 51, 1880.<br />
CARB 91<br />
Co-axial cellulose nan<strong>of</strong>ibers for electrical applications<br />
Minoru Miyauchi (1)(2) , minorumiyauchi@gmail.com, 110 8th Street, BT4123,<br />
Troy NY 12180, United States ; Jianjun Miao (1)(3)(4)(2) ; Trevor J. Simmons (1)(4)(2) ;<br />
Jonathan S. Dordick (1)(3)(5)(2) ; Robert J. Linhardt (1)(3)(4)(5)(2) . (1) Department <strong>of</strong><br />
Center for Nanotechnology, Rensselaer Polytechnic Institute, Troy NY 12180,<br />
United States (2) Department <strong>of</strong> Center for Biotechnology and Interdisciplinary<br />
Studies, Rensselaer Polytechnic Institute, Troy NY 12180, United States (3)<br />
Department <strong>of</strong> <strong>Chemical</strong> and Biological Engineering, Rensselaer Polytechnic<br />
Institute, Troy NY 12180, United States (4) Department <strong>of</strong> <strong>Chemistry</strong> and<br />
<strong>Chemical</strong> Biology, Rensselaer Polytechnic Institute, Troy NY 12180, United<br />
States (5) Department <strong>of</strong> Biology, Rensselaer Polytechnic Institute, Troy NY<br />
12180, United States<br />
Electrospinning has been widely investigated as simple technique to prepare<br />
nanoscale fibers <strong>of</strong> various polymers. New technologies are still being introduced<br />
for electrospinning, for example, dry-jet wet electrospinning using room<br />
temperature ionic liquids (RTILs) as solvent. Since RTILs are non-volatile,<br />
electrospun fibers are collected into a coagulation bath that is filled with<br />
appropriate co-solvent to remove RTILs. Because <strong>of</strong> the strong dissolving power<br />
<strong>of</strong> RTILs, electrospinning can be applied for many kinds <strong>of</strong> biomaterials, including<br />
unmodified cellulose. Some composite nan<strong>of</strong>ibers were fabricated by adding<br />
functional nanomaterials such as nanoparticles and carbon nanotubes into<br />
electrospinning solutions. Co-electrospinning techniques were also used to<br />
fabricate core-sheath type composite nan<strong>of</strong>ibers. Using these techniques,<br />
functional materials, which can normally not be electrospun, could be fabricated<br />
into fibers. Specialized cellulose nan<strong>of</strong>iber for electronic components and their<br />
potential applications will be discussed.<br />
CARB 92<br />
Study on the relative reactivity <strong>of</strong> glycosyl acceptors in the glycosylations<br />
<strong>of</strong><br />
2-Azido-2-deoxy-galactosides
Jane Kalikanda (1) , jkalika1@binghamton.edu, 4400 Vestal Parkway East,<br />
Binghamton NY 13902, United States ; Zhitao Li (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>,<br />
Binghamton University, Binghamton NY 13902, United States<br />
The<br />
reactivity <strong>of</strong> acceptors depends on the nucleophilicity <strong>of</strong> the hydroxyl groups<br />
in partially protected carbohydrate molecules that in turn depends on their<br />
nature, their spatial orientation, conformation <strong>of</strong> the sugar ring and also on<br />
the presence <strong>of</strong> other protecting groups in the molecule. A better understanding<br />
<strong>of</strong> the relative reactivity <strong>of</strong> glycosyl acceptors is very helpful for developing<br />
a more efficient oligosaccharide synthesis.<br />
However, it is <strong>of</strong>ten difficult to determine the relative reactivity <strong>of</strong><br />
acceptors with different protecting groups and from different sugars. We recently<br />
observed that the stereoselectivity <strong>of</strong> 2-azido-2-deoxy-galactoside donors is<br />
highly dependent on the reactivity <strong>of</strong> acceptors. The stereochemical outcome <strong>of</strong> a<br />
series <strong>of</strong><br />
glycosylation reactions was used to characterize a library <strong>of</strong> glycosyl<br />
acceptors and provided a useful way to quantify the reactivity <strong>of</strong> acceptors<br />
CARB 93<br />
Synthesis <strong>of</strong> tailored glycoconjugates for the precise detection <strong>of</strong><br />
pathogens<br />
Ashish A. Kulkarni (1) , ashishkulkarni30@gmail.com, 506, Riddle Road,, Apt.<br />
No. 41, Cincinnati Ohio 45220, United States ; Suri S. Iyer (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, University <strong>of</strong> Cincinnati, Cincinnati Ohio 45220, United States<br />
<strong>Carbohydrate</strong>-protein interactions play a very important role in a variety <strong>of</strong><br />
essential biological processes such as adhesion, cell-cell communication and<br />
organ differentiation. Several infectious agents use the cell surface<br />
carbohydrates to gain entry and infect the cells. We are harnessing this<br />
recognition capability and developing molecules that can be used as integral<br />
components <strong>of</strong> biosensors. This work is highly significant because carbohydrates<br />
are highly stable under a variety <strong>of</strong> conditions, do not suffer from lot-to-lot<br />
variation, can be synthesized in large quantities and can be adapted to any<br />
platform. Additionally, different strains, including newly emerging strains can be<br />
identified rapidly. In this regard, we have developed chemically defined<br />
glycoconjugates using a versatile modular approach for capturing toxins, viruses<br />
and bacteria. Specifically, we have synthesized carbohydrate based recognition<br />
elements and attached these molecules to a scaffold via flexible<br />
oligoethylene glycol linkers. The multivalent unit has been covalently linked to a<br />
fluorescent molecule or a biotin. The design and synthesis <strong>of</strong> these multivalent<br />
ligands, covalent linkage <strong>of</strong> the scaffold to the ligands and the reporter to the<br />
scaffold will be presented.
CARB 93<br />
Synthesis <strong>of</strong> tailored glycoconjugates for the precise detection <strong>of</strong><br />
pathogens<br />
Ashish A. Kulkarni (1) , ashishkulkarni30@gmail.com, 506, Riddle Road,, Apt.<br />
No. 41, Cincinnati Ohio 45220, United States ; Suri S. Iyer (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, University <strong>of</strong> Cincinnati, Cincinnati Ohio 45220, United States<br />
<strong>Carbohydrate</strong>-protein interactions play a very important role in a variety <strong>of</strong><br />
essential biological processes such as adhesion, cell-cell communication and<br />
organ differentiation. Several infectious agents use the cell surface<br />
carbohydrates to gain entry and infect the cells. We are harnessing this<br />
recognition capability and developing molecules that can be used as integral<br />
components <strong>of</strong> biosensors. This work is highly significant because carbohydrates<br />
are highly stable under a variety <strong>of</strong> conditions, do not suffer from lot-to-lot<br />
variation, can be synthesized in large quantities and can be adapted to any<br />
platform. Additionally, different strains, including newly emerging strains can be<br />
identified rapidly. In this regard, we have developed chemically defined<br />
glycoconjugates using a versatile modular approach for capturing toxins, viruses<br />
and bacteria. Specifically, we have synthesized carbohydrate based recognition<br />
elements and attached these molecules to a scaffold via flexible<br />
oligoethylene glycol linkers. The multivalent unit has been covalently linked to a<br />
fluorescent molecule or a biotin. The design and synthesis <strong>of</strong> these multivalent<br />
ligands, covalent linkage <strong>of</strong> the scaffold to the ligands and the reporter to the<br />
scaffold will be presented.<br />
CARB 94<br />
Structure-activity studies <strong>of</strong> synthetic glycophosphatidylinositol anchored<br />
proteins<br />
Carl V Christianson (1) , carl.christianson@mpikg.mpg.de, Free University Berlin,<br />
Arnimallee 22, Berlin Berlin 12101, Germany ; Peter H Seeberger (1) . (1)<br />
Department <strong>of</strong> Biomolecular Systems, Max Planck Institute <strong>of</strong> Colloids and<br />
Interfaces, Berlin 12101, Germany<br />
The structural complexity <strong>of</strong> glycophosphatidylinositol (GPI) anchors has<br />
hindered studies relating structural elements to specific biological roles. Access<br />
to defined GPI structures through total synthesis 1 facilitates the modification <strong>of</strong><br />
proteins via native chemical ligation (NCL) to probe the importance <strong>of</strong><br />
glycosylation and lipidation patterns <strong>of</strong> GPI anchors. 2 Here we present studies <strong>of</strong><br />
GFP modified with fully synthetic GPI anchors (Figure 1.) and other probes in<br />
order to gain a better understanding <strong>of</strong> the role that this posttranslational<br />
modification plays in<br />
biological processes.
1. Liu, X.; Kwon, Y. U.; Seeberger, P. H., Convergent synthesis <strong>of</strong> a fully lipidated<br />
glycosylphosphatidylinositol anchor <strong>of</strong> Plasmodium falciparum. J. Am. Chem.<br />
Soc. 2005, 127, (14), 5004-5.<br />
2. Becker,C. F.; Liu, X.; Olschewski, D.; Castelli, R.; Seidel, R.; Seeberger, P. H.,<br />
Semisynthesis <strong>of</strong> a glycosylphosphatidylinositol-anchored prion protein. Angew.<br />
Chem. Int. Ed. Engl. 2008, 47, (43), 8215-9.<br />
CARB 94<br />
Structure-activity studies <strong>of</strong> synthetic glycophosphatidylinositol anchored<br />
proteins<br />
Carl V Christianson (1) , carl.christianson@mpikg.mpg.de, Free University Berlin,<br />
Arnimallee 22, Berlin Berlin 12101, Germany ; Peter H Seeberger (1) . (1)<br />
Department <strong>of</strong> Biomolecular Systems, Max Planck Institute <strong>of</strong> Colloids and<br />
Interfaces, Berlin 12101, Germany<br />
The structural complexity <strong>of</strong> glycophosphatidylinositol (GPI) anchors has<br />
hindered studies relating structural elements to specific biological roles. Access<br />
to defined GPI structures through total synthesis 1 facilitates the modification <strong>of</strong><br />
proteins via native chemical ligation (NCL) to probe the importance <strong>of</strong><br />
glycosylation and lipidation patterns <strong>of</strong> GPI anchors. 2 Here we present studies <strong>of</strong><br />
GFP modified with fully synthetic GPI anchors (Figure 1.) and other probes in<br />
order to gain a better understanding <strong>of</strong> the role that this posttranslational<br />
modification plays in biological processes.
1. Liu, X.; Kwon, Y. U.; Seeberger, P. H., Convergent synthesis <strong>of</strong> a fully lipidated<br />
glycosylphosphatidylinositol anchor <strong>of</strong> Plasmodium falciparum. J. Am. Chem.<br />
Soc. 2005, 127, (14), 5004-5.<br />
2. Becker,C. F.; Liu, X.; Olschewski, D.; Castelli, R.; Seidel, R.; Seeberger, P. H.,<br />
Semisynthesis <strong>of</strong> a glycosylphosphatidylinositol-anchored prion protein. Angew.<br />
Chem. Int. Ed. Engl. 2008, 47, (43), 8215-9.<br />
CARB 95<br />
Chemoenzymatic synthesis <strong>of</strong> heparan sulfate<br />
Renpeng Liu (1) , renpengl@email.unc.edu, 303 Beard Hall, Chapel Hill 27599,<br />
United States ; Jian Liu (1) ; Yongmei Xu (1) . (1) Eshelman school <strong>of</strong> Pharmacy,<br />
UNC-Chapel Hill, Chapel Hill NC 27599, United States<br />
Heparan sulfate (HS) participates in a variety <strong>of</strong> biological functions. The specific<br />
functions <strong>of</strong> HS are governed by the unique distributions <strong>of</strong> sulfated saccharide<br />
sequences; however, chemical synthesis <strong>of</strong> structurally defined HS remains<br />
extremely challenging. Here, we describe a method for an enzyme-based de<br />
novo synthesis <strong>of</strong> HS oligosaccharides with defined structures. We use bacteria<br />
heparosan glycosyltransferases to build up the HS backbones and position the<br />
GlcNS residue by providing unnatural monosaccharide donor, UDP-Ntrifluoroacetylglucosamine.<br />
By using this approach, we have prepared four HS octasaccharides with defined<br />
distribution <strong>of</strong> GlcNS residues. By controlling the position <strong>of</strong> GlcNS residue,we<br />
are able to regulate the modification site <strong>of</strong> O-sulfotransferases and C5epimerase<br />
to prepare oligosaccharides with defined structures. Our method
could be used as a general approach for synthesizing structurally defined HS<br />
oligosaccharides that could aid in the discovery <strong>of</strong> novel HS-based therapeutic<br />
agent.<br />
CARB 96<br />
Synthesis <strong>of</strong> a fluorous-tagged disaccharide for the enzymatic preparation<br />
<strong>of</strong> heparin oligosaccharides<br />
Sayaka Masuko (1) , masuks@rpi.edu, 110 8th Street, Troy NY 12180, United<br />
States ; Smritilekha Bera (1) ; Robert J. Linhardt (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong><br />
and <strong>Chemical</strong> Biology, Rensselaer Polytechnic Institute, Troy NY 12180, United<br />
States<br />
Heparin is a highly sulfated glycosaminoglycan (GAG) that is extensively used in<br />
medical practice as an anticoagulant. The preparation <strong>of</strong> heparin and its<br />
oligosaccharides can be accomplished by synthetic and enzymatic methods.<br />
However, the purification <strong>of</strong> these modified oligosaccharide products from the<br />
reaction mixture is laborious and low-yielding. Therefore, a fluorous tag<br />
technique will greatly simplify purification procedures and structural<br />
characterization <strong>of</strong> the products. A heparin disaccharide with a fluorous affinity<br />
Froc tag can be chemically synthesized in a small number <strong>of</strong> steps. The Froctagged<br />
disaccharide will be placed on the reducing end to accept UDP-sugar<br />
building blocks in the enzymatic preparation <strong>of</strong> heparin and heparin<br />
oligosaccharides.<br />
CARB 96<br />
Synthesis <strong>of</strong> a fluorous-tagged disaccharide for the enzymatic preparation<br />
<strong>of</strong> heparin oligosaccharides<br />
Sayaka Masuko (1) , masuks@rpi.edu, 110 8th Street, Troy NY 12180, United<br />
States ; Smritilekha Bera (1) ; Robert J. Linhardt (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong><br />
and <strong>Chemical</strong> Biology, Rensselaer Polytechnic Institute, Troy NY 12180, United<br />
States<br />
Heparin is a highly sulfated glycosaminoglycan (GAG) that is extensively used in<br />
medical practice as an anticoagulant. The preparation <strong>of</strong> heparin and its<br />
oligosaccharides can be accomplished by synthetic and enzymatic methods.<br />
However, the purification <strong>of</strong> these modified oligosaccharide products from the<br />
reaction mixture is laborious and low-yielding. Therefore, a fluorous tag<br />
technique will greatly simplify purification procedures and structural<br />
characterization <strong>of</strong> the products. A heparin disaccharide with a fluorous affinity<br />
Froc tag can be chemically synthesized in a small number <strong>of</strong> steps. The Froctagged<br />
disaccharide will be placed on the reducing end to accept UDP-sugar
uilding blocks in the enzymatic preparation <strong>of</strong> heparin and heparin<br />
oligosaccharides.<br />
CARB 97<br />
Preventing the transmission <strong>of</strong> Plasmodium falciparum through the<br />
inhibition <strong>of</strong> malaria protein binding to placental chondroitin sulfate A<br />
Julie M Beaudet (1)(2) , beaudj3@rpi.edu, 110 8th Street, Troy New York, United<br />
States ; Leandra J Mansur (1)(2) ; Bo Yang (1) ; Fuming Zhang (3) ; Robert J<br />
Linhardt (1)(2)(3) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer<br />
Polytechnic Institute, United States (2) Department <strong>of</strong> Biology, Rensselaer<br />
Polytechnic Institute, United States (3) Department <strong>of</strong> <strong>Chemical</strong> and Biological<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States<br />
One way that Plasmodium falciparum can be transferred is during the birthing<br />
process. Infected erythrocytes express the VAR2CSA protein which binds to<br />
chondroitin sulfate A (CSA) in the placenta. During birth the bound erythrocytes<br />
rupture and the P. falciparum parasite can infect the newborn. Laboratory studies<br />
have already shown that bovine tracheal CSA can inhibit the binding <strong>of</strong> infected<br />
erythrocytes to CSA in the placenta. We are isolating bovine placental<br />
glycosaminoglycans (GAGs), and can inhibit the infected cells from binding to the<br />
placenta using the extracted GAGs. The GAG -bound infected cells can be<br />
transported to the spleen thus preventing contact between the parasite and the<br />
newborn. The GAGs are isolated from the placenta tissue and purified. Structural<br />
analysis <strong>of</strong> the GAGs is preformed using PAGE, NMR, and LCMS disaccharide<br />
analysis techniques. Extracted CSA is then immobilized on an SPR chip in order<br />
to perform an activity assay with infected erythrocytes or similar protein<br />
compounds.<br />
CARB 97<br />
Preventing the transmission <strong>of</strong> Plasmodium falciparum through the<br />
inhibition <strong>of</strong> malaria protein binding to placental chondroitin sulfate A<br />
Julie M Beaudet (1)(2) , beaudj3@rpi.edu, 110 8th Street, Troy New York, United<br />
States ; Leandra J Mansur (1)(2) ; Bo Yang (1) ; Fuming Zhang (3) ; Robert J<br />
Linhardt (1)(2)(3) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer<br />
Polytechnic Institute, United States (2) Department <strong>of</strong> Biology, Rensselaer<br />
Polytechnic Institute, United States (3) Department <strong>of</strong> <strong>Chemical</strong> and Biological<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States<br />
One way that Plasmodium falciparum can be transferred is during the birthing<br />
process. Infected erythrocytes express the VAR2CSA protein which binds to
chondroitin sulfate A (CSA) in the placenta. During birth the bound erythrocytes<br />
rupture and the P. falciparum parasite can infect the newborn. Laboratory studies<br />
have already shown that bovine tracheal CSA can inhibit the binding <strong>of</strong> infected<br />
erythrocytes to CSA in the placenta. We are isolating bovine placental<br />
glycosaminoglycans (GAGs), and can inhibit the infected cells from binding to the<br />
placenta using the extracted GAGs. The GAG -bound infected cells can be<br />
transported to the spleen thus preventing contact between the parasite and the<br />
newborn. The GAGs are isolated from the placenta tissue and purified. Structural<br />
analysis <strong>of</strong> the GAGs is preformed using PAGE, NMR, and LCMS disaccharide<br />
analysis techniques. Extracted CSA is then immobilized on an SPR chip in order<br />
to perform an activity assay with infected erythrocytes or similar protein<br />
compounds.<br />
CARB 98<br />
Real-time assessment <strong>of</strong> the morphological change <strong>of</strong> cellulose in<br />
response to enzymatic treatment<br />
Chi Nguyen (1) , cn382@drexel.edu, 3141 Chestnut Street, Philadelphia Pa<br />
19104, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong>, Drexel University,<br />
Philadelphia Pa 19103, United States<br />
Cellulose is considered a very attractive alternative source <strong>of</strong> fuels because <strong>of</strong><br />
the abundant and renewable supply. Thus improving conversion <strong>of</strong> cellulose from<br />
biomass into cellulosic ethanol is <strong>of</strong> great scientific interest. Presently, one <strong>of</strong> the<br />
key steps <strong>of</strong> the enzymatic hydrolysis <strong>of</strong> cellulose, where cellulase breaks down<br />
the crystal structure on the surface region <strong>of</strong> cellulose and exposes cellulose<br />
chains for subsequent hydrolysis by cellulase, is still not fully understood. It is<br />
hypothesized that this non-hydrolytic enzymatic step could be the rate-limiting<br />
step for the entire enzymatic hydrolysis <strong>of</strong> cellulose. We have developed a<br />
nanomechanical approach that allows us to assess the minute change in<br />
morphology <strong>of</strong> cellulose in real time. Here we report our recent progress in using<br />
this approach to study the effect <strong>of</strong> cellulolytic enzyme on cellulose. The results<br />
<strong>of</strong> this study will help to provide a thorough insight into the initial steps <strong>of</strong><br />
cellulose hydrolysis.<br />
CARB 98<br />
Real-time assessment <strong>of</strong> the morphological change <strong>of</strong> cellulose in<br />
response to enzymatic treatment<br />
Chi Nguyen (1) , cn382@drexel.edu, 3141 Chestnut Street, Philadelphia Pa<br />
19104, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong>, Drexel University,<br />
Philadelphia Pa 19103, United States
Cellulose is considered a very attractive alternative source <strong>of</strong> fuels because <strong>of</strong><br />
the abundant and renewable supply. Thus improving conversion <strong>of</strong> cellulose from<br />
biomass into cellulosic ethanol is <strong>of</strong> great scientific interest. Presently, one <strong>of</strong> the<br />
key steps <strong>of</strong> the enzymatic hydrolysis <strong>of</strong> cellulose, where cellulase breaks down<br />
the crystal structure on the surface region <strong>of</strong> cellulose and exposes cellulose<br />
chains for subsequent hydrolysis by cellulase, is still not fully understood. It is<br />
hypothesized that this non-hydrolytic enzymatic step could be the rate-limiting<br />
step for the entire enzymatic hydrolysis <strong>of</strong> cellulose. We have developed a<br />
nanomechanical approach that allows us to assess the minute change in<br />
morphology <strong>of</strong> cellulose in real time. Here we report our recent progress in using<br />
this approach to study the effect <strong>of</strong> cellulolytic enzyme on cellulose. The results<br />
<strong>of</strong> this study will help to provide a thorough insight into the initial steps <strong>of</strong><br />
cellulose hydrolysis.<br />
CARB 99<br />
Chemoselective glycosylation <strong>of</strong> hemoglobin as a potential oxygen<br />
therapeutic<br />
Thomas J Styslinger (1) , styslinger.1@osu.edu, Newman & Wolfrom Lab, 100 W.<br />
18th Avenue, Columbus Ohio 43210, United States ; Ning Zhang (2) ; Andre F.<br />
Palmer (2) ; Peng G. Wang (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>, The Ohio State<br />
University, Columbus Ohio 43210, United States (2) Department <strong>of</strong> <strong>Chemical</strong> and<br />
Biomolecular Engineering, The Ohio State University, Columbus Ohio 43210,<br />
United States<br />
Hemorrhagic shock is the leading cause <strong>of</strong> death among trauma patients 1 . This<br />
condition results from the body's inability to compensate and provide adequate<br />
tissue perfusion and oxygenation as a result <strong>of</strong> blood loss. Over the past 20<br />
years, hemoglobin-based oxygen carriers (HBOCs) have been explored as a<br />
potential treatment for hemorrhagic shock due to hemoglobin's role in the<br />
transportation and storage <strong>of</strong> oxygen in red blood cells. One attractive potential<br />
for the development <strong>of</strong> HBOCs involves taking advantage <strong>of</strong> nature's strategy <strong>of</strong><br />
protein glycosylation for the protection and stabilization <strong>of</strong> proteins. Previous<br />
studies conducted in our labs have shown that site-selective glycosylation <strong>of</strong><br />
hemoglobin is possible with a lactose derivative and has advantageous effects 2 .<br />
In the present study we have expanded the glycosylation sites to include lysine<br />
residues and developed methodology to facilely broaden the scope <strong>of</strong><br />
carbohydrates conjugated to hemoglobin. Furthermore, the synthetic route<br />
employed avoids the tedious protection and deprotection steps which typically<br />
accompany the synthesis <strong>of</strong> such molecules. The range <strong>of</strong> carbohydrates being<br />
investigated for conjugation and corresponding biological studies includes<br />
various oligosaccharides and polysaccharides.<br />
1) Cocchi, M. N., Kimlin, E., Walsh, M., Donnino, M. W., Emerg Med Clin North<br />
Am. 2007, 25(3), 623-42.
2) Zhang, Y., Bhatt, V. S., Sun, G., Wang, P. G., Palmer, A. F., Bioconjugat<br />
Chem. 2008, 19, 2221-2230.<br />
CARB 100<br />
Immobilization <strong>of</strong> enzymes relevant to bioengineered heparin synthesis<br />
Eric R. Sterner (1) , sterne@rpi.edu, 110 8th Street, Center for Biotechnology and<br />
Interdisciplinary Studies, Troy New York 12180, United States ; Robert J.<br />
Linhardt (2) ; Jonathan S. Dordick (1) ; Jian Liu (3) ; Fuming Zhang (2) ; Wenjing Zhao (4) ;<br />
Priscilla Paul (1) ; Jeff Martin (5) . (1) Department <strong>of</strong> <strong>Chemical</strong> and Biochemical<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States (2) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy New York 12180, United States (3) Department <strong>of</strong><br />
Medicinal <strong>Chemistry</strong>, University <strong>of</strong> North Carolina at Chapel Hill, Chapel Hill<br />
North Carolina 27599, United States (4) Department <strong>of</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy New York 12180, United States (5) Department <strong>of</strong><br />
Biochemistry and Biophysics, Rensselaer Polytechnic Institute, Troy New York<br />
12180, United States<br />
The immobilization <strong>of</strong> enzymes on solid resin surfaces is an important technique<br />
used as a means to improve the stability and performance <strong>of</strong> enzymes for a<br />
variety <strong>of</strong> applications, as well as a method to eliminate costly downstream<br />
separations steps required for free enzymes in solution. This study aims to<br />
investigate the expression, purification, and immobilization <strong>of</strong> the Osulfotransferase<br />
enzymes relevant to the synthesis <strong>of</strong> bioengineered heparin.<br />
Enzyme immobilization will be performed on micro-particle with intended<br />
applications in the medical and biochemistry research and process scale-up.<br />
Immobilization on nano-particles will also be considered with applications<br />
intended for heparin synthesis on a micro-scale. The kinetics <strong>of</strong> the free and<br />
immobilized enzymes will be determined and compared to illustrate the benefit <strong>of</strong><br />
immobilized enzymes reactions over free enzyme solutions. A thorough study <strong>of</strong><br />
the effect <strong>of</strong> solid resin affinity immobilization on enzyme stability will also be<br />
considered and reported.<br />
CARB 100<br />
Immobilization <strong>of</strong> enzymes relevant to bioengineered heparin synthesis<br />
Eric R. Sterner (1) , sterne@rpi.edu, 110 8th Street, Center for Biotechnology and<br />
Interdisciplinary Studies, Troy New York 12180, United States ; Robert J.<br />
Linhardt (2) ; Jonathan S. Dordick (1) ; Jian Liu (3) ; Fuming Zhang (2) ; Wenjing Zhao (4) ;<br />
Priscilla Paul (1) ; Jeff Martin (5) . (1) Department <strong>of</strong> <strong>Chemical</strong> and Biochemical<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States (2) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer
Polytechnic Institute, Troy New York 12180, United States (3) Department <strong>of</strong><br />
Medicinal <strong>Chemistry</strong>, University <strong>of</strong> North Carolina at Chapel Hill, Chapel Hill<br />
North Carolina 27599, United States (4) Department <strong>of</strong> Biology, Rensselaer<br />
Polytechnic Institute, Troy New York 12180, United States (5) Department <strong>of</strong><br />
Biochemistry and Biophysics, Rensselaer Polytechnic Institute, Troy New York<br />
12180, United States<br />
The immobilization <strong>of</strong> enzymes on solid resin surfaces is an important technique<br />
used as a means to improve the stability and performance <strong>of</strong> enzymes for a<br />
variety <strong>of</strong> applications, as well as a method to eliminate costly downstream<br />
separations steps required for free enzymes in solution. This study aims to<br />
investigate the expression, purification, and immobilization <strong>of</strong> the Osulfotransferase<br />
enzymes relevant to the synthesis <strong>of</strong> bioengineered heparin.<br />
Enzyme immobilization will be performed on micro-particle with intended<br />
applications in the medical and biochemistry research and process scale-up.<br />
Immobilization on nano-particles will also be considered with applications<br />
intended for heparin synthesis on a micro-scale. The kinetics <strong>of</strong> the free and<br />
immobilized enzymes will be determined and compared to illustrate the benefit <strong>of</strong><br />
immobilized enzymes reactions over free enzyme solutions. A thorough study <strong>of</strong><br />
the effect <strong>of</strong> solid resin affinity immobilization on enzyme stability will also be<br />
considered and reported.<br />
CARB 101<br />
Analyses <strong>of</strong> anti Tn-antigen MLS128 monoclonal antibody binding to two or<br />
three consecutive Tn-antigen clusters by surface plasmon resonance (SPR)<br />
and NMR<br />
Ayano Takasaki-Matsumoto (1) , ayanot@keyaki.cc.u-tokai.ac.jp, 4-1-1<br />
Kitakaname, Hiratsuka Kanagawa 259-1292, Japan ; Shinya Hanashima (2) ; Ami<br />
Aoki (1) ; Yoshiki Yamaguchi (2) ; Reiko Sato (3) ; Hiroko Kawakami (3) ; Mamoru<br />
Mizuno (3) ; Yoko Fujita-Yamaguchi (1) . (1) Department <strong>of</strong> Applied Biochemistry,<br />
Tokai University School <strong>of</strong> Engineering, Hiratsuka Kanagawa 259-1292, Japan<br />
(2) Frontier Research System, RIKEN, Wako Saitama 351-0198, Japan (3)<br />
Glyco-organic <strong>Chemistry</strong>, The Noguchi Institute, Itabashi Tokyo 173-0003, Japan<br />
Tn-antigens (GalNAca-Ser/Thr) are oncogenic antigens and are<br />
generally masked by covalently linked terminal carbohydrate moieties in normal<br />
human tissues but are exposed in most primary and metastatic epithelial<br />
malignant tumors. MLS128 recongnizes an epitope consisting <strong>of</strong> three or two<br />
consecutive Tn-antigens (Tn3 or Tn2). Thermodynamic and kinetic analyses <strong>of</strong><br />
the antigen-antibody interaction between MLS128 and Tn3- or Tn2-peptide,<br />
synthesized as described (Sakai et al. J. Biochem. 2010), were conducted by<br />
SPR and NMR at varying temperatures. The binding epitopes were analyzed by<br />
1 H-NMR as well as TR-NOESY experiments. At varying temperatures from 5 to<br />
35 °C, KD values for both ligands increased while affinities for Tn3 were always
higher than Tn2 as determined by SPR. NMR experiments supported that the<br />
ligand binding site <strong>of</strong> MLS128 accepts three GalNAc residues on Tn3-peptide,<br />
which correlated with the SPR results that the affinity for Tn3-peptide exhibited<br />
lower KD value than that <strong>of</strong> Tn3-peptide.<br />
CARB 101<br />
Analyses <strong>of</strong> anti Tn-antigen MLS128 monoclonal antibody binding to two or<br />
three consecutive Tn-antigen clusters by surface plasmon resonance (SPR)<br />
and NMR<br />
Ayano Takasaki-Matsumoto (1) , ayanot@keyaki.cc.u-tokai.ac.jp, 4-1-1<br />
Kitakaname, Hiratsuka Kanagawa 259-1292, Japan ; Shinya Hanashima (2) ; Ami<br />
Aoki (1) ; Yoshiki Yamaguchi (2) ; Reiko Sato (3) ; Hiroko Kawakami (3) ; Mamoru<br />
Mizuno (3) ; Yoko Fujita-Yamaguchi (1) . (1) Department <strong>of</strong> Applied Biochemistry,<br />
Tokai University School <strong>of</strong> Engineering, Hiratsuka Kanagawa 259-1292, Japan<br />
(2) Frontier Research System, RIKEN, Wako Saitama 351-0198, Japan (3)<br />
Glyco-organic <strong>Chemistry</strong>, The Noguchi Institute, Itabashi Tokyo 173-0003, Japan<br />
Tn-antigens (GalNAca-Ser/Thr) are oncogenic antigens and are<br />
generally masked by covalently linked terminal carbohydrate moieties in normal<br />
human tissues but are exposed in most primary and metastatic epithelial<br />
malignant tumors. MLS128 recongnizes an epitope consisting <strong>of</strong> three or two<br />
consecutive Tn-antigens (Tn3 or Tn2). Thermodynamic and kinetic analyses <strong>of</strong><br />
the antigen-antibody interaction between MLS128 and Tn3- or Tn2-peptide,<br />
synthesized as described (Sakai et al. J. Biochem. 2010), were conducted by<br />
SPR and NMR at varying temperatures. The binding epitopes were analyzed by<br />
1 H-NMR as well as TR-NOESY experiments. At varying temperatures from 5 to<br />
35 °C, KD values for both ligands increased while affinities for Tn3 were always<br />
higher than Tn2 as determined by SPR. NMR experiments supported that the<br />
ligand binding site <strong>of</strong> MLS128 accepts three GalNAc residues on Tn3-peptide,<br />
which correlated with the SPR results that the affinity for Tn3-peptide exhibited<br />
lower KD value than that <strong>of</strong> Tn3-peptide.<br />
CARB 102<br />
Modular glycoconjugate tool set for assembly and presentation <strong>of</strong><br />
multivalent carbohydrate ligands on surfaces<br />
Irene Abia (1) , dcbaker@utk.edu, 1420 Circle Drive, Knoxville TN 37996-1600,<br />
United States ; Brian Sanders (1) ; Michael D. Best (1) ; David C. Baker (1) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong>, University <strong>of</strong> Tennessee, Knoxville TN 37996-1600,<br />
United States<br />
D-Mannose mono-, di- and trisaccharides with thiol-terminated aglycons were<br />
synthesized for attachment to gold nanoparticles to mimic carbohydrate-involved
cell-surface interactions. These molecules were constructed by glycosylation <strong>of</strong><br />
appropriately protected glycosyl donors and acceptors, followed by free-radical<br />
addition to introduce the thiol terminals onto the aglycons. Subsequent<br />
deprotection afforded the free-OH saccharides. Tethering chemistries will allow<br />
these components to be attached to gold-clad silicon chips, creating<br />
carbohydrate surfaces <strong>of</strong> varying valencies and densities. Propargyl glycosides<br />
<strong>of</strong> mono- and di-mannosides have also been synthesized for tethering via click<br />
chemistry. (Supported by NSF DMR-0906752.)<br />
CARB 102<br />
Modular glycoconjugate tool set for assembly and presentation <strong>of</strong><br />
multivalent carbohydrate ligands on surfaces<br />
Irene Abia (1) , dcbaker@utk.edu, 1420 Circle Drive, Knoxville TN 37996-1600,<br />
United States ; Brian Sanders (1) ; Michael D. Best (1) ; David C. Baker (1) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong>, University <strong>of</strong> Tennessee, Knoxville TN 37996-1600,<br />
United States<br />
D-Mannose mono-, di- and trisaccharides with thiol-terminated aglycons were<br />
synthesized for attachment to gold nanoparticles to mimic carbohydrate-involved<br />
cell-surface interactions. These molecules were constructed by glycosylation <strong>of</strong><br />
appropriately protected glycosyl donors and acceptors, followed by free-radical<br />
addition to introduce the thiol terminals onto the aglycons. Subsequent<br />
deprotection afforded the free-OH saccharides. Tethering chemistries will allow<br />
these components to be attached to gold-clad silicon chips, creating<br />
carbohydrate surfaces <strong>of</strong> varying valencies and densities. Propargyl glycosides<br />
<strong>of</strong> mono- and di-mannosides have also been synthesized for tethering via click<br />
chemistry. (Supported by NSF DMR-0906752.)<br />
CARB 103<br />
Analysis <strong>of</strong> the absorption <strong>of</strong> low molecular weight heparin in human<br />
umbilical cord tissue as a model for the prevention <strong>of</strong> cancer metastasis
Amanda M. Weyers (1) , weyera@rpi.edu, 110 8th St., Cogswell, Troy NY 12180,<br />
United States ; Thangriala Sudha (2) ; Bo Yang (1) ; Boyangzi Li (1) ; Majde<br />
Takieddin (2) ; Fuming Zhang (3) ; Shaker A. Mousa (2) ; Robert J. Linhardt (1)(4) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic<br />
Institute, Troy NY 12180, United States (2) The Pharmaceutical Research<br />
Institute, Albany College <strong>of</strong> Pharmacy and Health Sciences, Albany NY 12208,<br />
United States (3) Department <strong>of</strong> <strong>Chemical</strong> and Biological Engineering,<br />
Rensselaer Polytechnic Institute, Troy NY 12180, United States (4) Center for<br />
Biotechnology and Interdisciplinary Studies, Rensselaer Polytechnic Institute,<br />
Troy NY 12180, United States<br />
Glycosoaminoglycans (GAGs) mediate a wide variety <strong>of</strong> biological functions. Low<br />
molecular weight heparins (LMWHs) re GAG derivatives exhibiting favorable<br />
pharmacological properties are being studied as anti-cancer drugs. Human<br />
pancreatic cancer cell adhesion has been shown to be inhibited by LMWH,<br />
possibly via the binding <strong>of</strong> GAG to the endothelial lining <strong>of</strong> the blood vessel. Here<br />
a non-anticoagulant LMWH was perfused through the vein in the umbilical cord<br />
tissue samples at different concentrations to determine the absorption <strong>of</strong> GAG by<br />
the blood vessel. Samples were then defatted, proteolyzed, purified by anionexchange<br />
chromatography, and subject to methanol precipitation. Total sulfated<br />
GAG content <strong>of</strong> the isolated samples was quantified via carbazole assay and<br />
through complexation with 9-dimethymethylene blue. The polydispersity and<br />
molecular weights <strong>of</strong> the isolates was determined via polyacrylamide gel<br />
electrophoresis and GAG structures were determined via liquid chromatographymass<br />
spectroscopy.<br />
CARB 104<br />
Synthesis and biological evaluation <strong>of</strong> a Gal(α1-2)GalCer analog<br />
Yanke Liang (1) , yanke.liang@uconn.edu, 55 N. Eagleville Road, Storrs CT<br />
06269-3060, United States ; Amy R Howell (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>,<br />
University <strong>of</strong> Connecticut, Storrs CT 06269-3060, United States
A synthesis <strong>of</strong> an analog <strong>of</strong> the immunostimulatory glycolipid, Gal(α1-<br />
2)galactosylceramide, is described. Good reactivity and high α selectivity was<br />
achieved utilizing 'armed' donors and 'disarmed' acceptors. Biological evaluation<br />
is expected to establish the effect <strong>of</strong> antigen presenting cells on the cytokine<br />
release pr<strong>of</strong>ile.<br />
CARB 104<br />
Synthesis and biological evaluation <strong>of</strong> a Gal(α1-2)GalCer analog<br />
Yanke Liang (1) , yanke.liang@uconn.edu, 55 N. Eagleville Road, Storrs CT<br />
06269-3060, United States ; Amy R Howell (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>,<br />
University <strong>of</strong> Connecticut, Storrs CT 06269-3060, United States<br />
A synthesis <strong>of</strong> an analog <strong>of</strong> the immunostimulatory glycolipid, Gal(α1-<br />
2)galactosylceramide, is described. Good reactivity and high α selectivity was<br />
achieved utilizing 'armed' donors and 'disarmed' acceptors. Biological evaluation<br />
is expected to establish the effect <strong>of</strong> antigen presenting cells on the cytokine<br />
release pr<strong>of</strong>ile.<br />
CARB 105<br />
Thiol-click chemistry approach to glycomimetics: Novel stereoselective<br />
synthesis <strong>of</strong><br />
(1-3)-S-thiodisaccharides<br />
Irena Bak-Sypien (1) , 84 W. South Street, Wilkes-Barre PA 18766, United States ;<br />
Zbigniew J. Witczak (1) . (1) Department <strong>of</strong> Pharmaceutical Sciences, Wilkes<br />
University, Wilkes-Barre PA 18766, United States<br />
During the last decade the concept <strong>of</strong> click chemistry was transplanted to<br />
carbohydrate chemistry [1-3]. While, the original alkyne-azide concept <strong>of</strong> click-<br />
chemistry is well known, the alternative thiol-click is relatively unexplored.<br />
Recently, we have developed one-pot, thiol-click procedure for the synthesis <strong>of</strong><br />
new family <strong>of</strong> (1-5)-C-thiodisaccharides [3]. Starting template for the specifically<br />
developed thiol-click approach is cyanoglycal 1 conveniently prepared in our
laboratory.<br />
Glycal 1 undergoes base catalyzed Michael additions with peracetylated 1-thio-<br />
D-glucose 2 via one step thiol-click approach. The exclusive regio- and<br />
stereochemistry <strong>of</strong> the Michael addition reaction proceeds with the formation <strong>of</strong><br />
1,4-adduct 3 in high yield. The adduct 3 upon the conventional mild hydrolysis,<br />
affords amide derivative suitable for further functionalization with protected<br />
aminoacids to form carboamino peptides. Further examples <strong>of</strong> reactive thiols for<br />
quick one-pot thiol-click coupling approach with functionalized enones and their<br />
application to synthesis <strong>of</strong> new families <strong>of</strong> thio carboamino peptides will be also<br />
explored. They will be use as new tools for glycobiology and specifically as new<br />
inhibitors <strong>of</strong> galectin-3 [4].<br />
1. M.Fiore, A. Mara, and A. Dondoni, J. Org.. Chem., 2009, 74, 4222.<br />
2. S. G. Gouin, L. Bultel, C. Falentin and J. Kovensky, Eur. J. Org. Chem. 2007,<br />
1160.<br />
3. Z. J. Witczak, D. Lorchak and N. Nguyen, Carbohydr. Res., 2007, 342, 1929.<br />
4. Galectins, John Wiley & Sons, Hoboken, NJ, 2008, Eds. A. Klyosov, Z.J.<br />
Witczak, and D. Platt.<br />
CARB 105<br />
Thiol-click chemistry approach to glycomimetics: Novel stereoselective<br />
synthesis <strong>of</strong><br />
(1-3)-S-thiodisaccharides<br />
Irena Bak-Sypien (1) , 84 W. South Street, Wilkes-Barre PA 18766, United States ;<br />
Zbigniew J. Witczak (1) . (1) Department <strong>of</strong> Pharmaceutical Sciences, Wilkes<br />
University, Wilkes-Barre PA 18766, United States<br />
During the last decade the concept <strong>of</strong> click chemistry was transplanted to<br />
carbohydrate chemistry [1-3]. While, the original alkyne-azide concept <strong>of</strong> click-<br />
chemistry is well known, the alternative thiol-click is relatively unexplored.<br />
Recently, we have developed one-pot, thiol-click procedure for the synthesis <strong>of</strong><br />
new family <strong>of</strong> (1-5)-C-thiodisaccharides [3]. Starting template for the specifically
developed thiol-click approach is cyanoglycal 1 conveniently prepared in our<br />
laboratory.<br />
Glycal 1 undergoes base catalyzed Michael additions with peracetylated 1-thio-<br />
D-glucose 2 via one step thiol-click approach. The exclusive regio- and<br />
stereochemistry <strong>of</strong> the Michael addition reaction proceeds with the formation <strong>of</strong><br />
1,4-adduct 3 in high yield. The adduct 3 upon the conventional mild hydrolysis,<br />
affords amide derivative suitable for further functionalization with protected<br />
aminoacids to form carboamino peptides. Further examples <strong>of</strong> reactive thiols for<br />
quick one-pot thiol-click coupling approach with functionalized enones and their<br />
application to synthesis <strong>of</strong> new families <strong>of</strong> thio carboamino peptides will be also<br />
explored. They will be use as new tools for glycobiology and specifically as new<br />
inhibitors <strong>of</strong> galectin-3 [4].<br />
1. M.Fiore, A. Mara, and A. Dondoni, J. Org.. Chem., 2009, 74, 4222.<br />
2. S. G. Gouin, L. Bultel, C. Falentin and J. Kovensky, Eur. J. Org. Chem. 2007,<br />
1160.<br />
3. Z. J. Witczak, D. Lorchak and N. Nguyen, Carbohydr. Res., 2007, 342, 1929.<br />
4. Galectins, John Wiley & Sons, Hoboken, NJ, 2008, Eds. A. Klyosov, Z.J.<br />
Witczak, and D. Platt.<br />
CARB 106<br />
High-throughput glycoarray for monitoring immune responses to a cancer<br />
vaccine<br />
Christopher Campbell (1) , campbellct@mail.nih.gov, 376 Boyles Street, Room<br />
108, Frederick MD 21702, United States ; Yalong Zhang (1) ; Olaf Ludek (1) ; David<br />
Farnsworth (1) ; Jeffrey Gildersleeve (1) . (1) <strong>Chemical</strong> Biology Laboratory, National<br />
Cancer Institute, Frederick MD 21702, United States<br />
Cancer vaccines lengthen survival in some patients, though others experience<br />
little or no benefit. Better understanding <strong>of</strong> tumor characteristics and host factors<br />
that influence the response to vaccination should help explain these survival<br />
differences between patients and lead to more widely effective vaccines.
Important insights will likely come from monitoring humoral responses to<br />
carbohydrates. Although ubiquitious in cancer, abnormal tumor-associated<br />
carbohydrate antigens are overlooked aspects <strong>of</strong> antitumor immunity due to<br />
technical challenges <strong>of</strong> studying carbohydrate-protein interactions. We used a<br />
high-throughput glycoarray to study humoral responses induced in 28 subjects by<br />
a prostate cancer vaccine (PSA TRICOM). Post-vaccination changes in anticarbohydrate<br />
antibodies were detected in the majority <strong>of</strong> patients, including<br />
responses to known tumor-associated carbohydrate antigens and the terminal<br />
disaccharide <strong>of</strong> the Forssman antigen (GalNAcα1-3GalNAcβ). These changes<br />
might reflect reactions to vaccine components or antigen spreading. Studies<br />
aimed at elucidating the origin <strong>of</strong> these changes are ongoing.<br />
CARB 106<br />
High-throughput glycoarray for monitoring immune responses to a cancer<br />
vaccine<br />
Christopher Campbell (1) , campbellct@mail.nih.gov, 376 Boyles Street, Room<br />
108, Frederick MD 21702, United States ; Yalong Zhang (1) ; Olaf Ludek (1) ; David<br />
Farnsworth (1) ; Jeffrey Gildersleeve (1) . (1) <strong>Chemical</strong> Biology Laboratory, National<br />
Cancer Institute, Frederick MD 21702, United States<br />
Cancer vaccines lengthen survival in some patients, though others experience<br />
little or no benefit. Better understanding <strong>of</strong> tumor characteristics and host factors<br />
that influence the response to vaccination should help explain these survival<br />
differences between patients and lead to more widely effective vaccines.<br />
Important insights will likely come from monitoring humoral responses to<br />
carbohydrates. Although ubiquitious in cancer, abnormal tumor-associated<br />
carbohydrate antigens are overlooked aspects <strong>of</strong> antitumor immunity due to<br />
technical challenges <strong>of</strong> studying carbohydrate-protein interactions. We used a<br />
high-throughput glycoarray to study humoral responses induced in 28 subjects by<br />
a prostate cancer vaccine (PSA TRICOM). Post-vaccination changes in anticarbohydrate<br />
antibodies were detected in the majority <strong>of</strong> patients, including<br />
responses to known tumor-associated carbohydrate antigens and the terminal<br />
disaccharide <strong>of</strong> the Forssman antigen (GalNAcα1-3GalNAcβ). These changes<br />
might reflect reactions to vaccine components or antigen spreading. Studies<br />
aimed at elucidating the origin <strong>of</strong> these changes are ongoing.<br />
CARB 107<br />
NMR spectroscopic studies <strong>of</strong> APF: A small glycopeptide possessing<br />
potent antiproliferative activity<br />
Kristie M Adams (1) , adamskm@mail.nih.gov, 376 Boyles St, Bldg 376, Frederick<br />
Maryland 21702, United States ; Piotr Kaczmarek (1) ; Susan K Keay (2)(3) ; Joseph J<br />
Barchi, Jr (1) . (1) <strong>Chemical</strong> Biology Laboratory, National Cancer Institute-
Frederick, Frederick Maryland 21702, United States (2) Department <strong>of</strong> Medicine,<br />
<strong>Division</strong> <strong>of</strong> Infectious Diseases, University <strong>of</strong> Maryland School <strong>of</strong> Medicine,<br />
Baltimore Maryland 21201, United States (3) Research Service, Veterans Affairs<br />
Maryland Health Care System, Baltimore Maryland 21201, United States<br />
Antiproliferative factor (APF) is a novel glycopeptide isolated from the urine <strong>of</strong><br />
patients diagnosed with interstitial cystitis/painful bladder syndrome (IC/PBS), a<br />
chronic bladder disease. APF is a negative growth factor that inhibits proliferation<br />
<strong>of</strong> normal bladder epithelial cells at subnanomolar concentrations, leading to the<br />
hypothesis that APF could possess anticancer properties. APF was found to<br />
markedly inhibit growth <strong>of</strong> both T24 bladder carcinoma and kidney cancer cell<br />
lines. Subsequent structural and SAR studies revealed that APF (as isolated<br />
from IC/PBS patients) is a nine-residue peptide containing sialyl-TF antigen α-Olinked<br />
to the N-terminal threonine residue, Neu5Acα2-3Galβ1-3GalNAcα-O-<br />
TVPAAVVVA. Neither the sialyl group nor the C-terminal alanine residue is<br />
necessary to maintain antiproliferative activity, yet other seemingly<br />
inconsequential structural changes completely abolish the activity <strong>of</strong> APF. Thus,<br />
we have structurally characterized APF and several APF analogues in aqueous<br />
solution using 2D NMR spectroscopic techniques in an attempt to further<br />
understand the structure <strong>of</strong> APF.<br />
CARB 107<br />
NMR spectroscopic studies <strong>of</strong> APF: A small glycopeptide possessing<br />
potent antiproliferative activity<br />
Kristie M Adams (1) , adamskm@mail.nih.gov, 376 Boyles St, Bldg 376, Frederick<br />
Maryland 21702, United States ; Piotr Kaczmarek (1) ; Susan K Keay (2)(3) ; Joseph J<br />
Barchi, Jr (1) . (1) <strong>Chemical</strong> Biology Laboratory, National Cancer Institute-<br />
Frederick, Frederick Maryland 21702, United States (2) Department <strong>of</strong> Medicine,<br />
<strong>Division</strong> <strong>of</strong> Infectious Diseases, University <strong>of</strong> Maryland School <strong>of</strong> Medicine,<br />
Baltimore Maryland 21201, United States (3) Research Service, Veterans Affairs<br />
Maryland Health Care System, Baltimore Maryland 21201, United States<br />
Antiproliferative factor (APF) is a novel glycopeptide isolated from the urine <strong>of</strong><br />
patients diagnosed with interstitial cystitis/painful bladder syndrome (IC/PBS), a<br />
chronic bladder disease. APF is a negative growth factor that inhibits proliferation<br />
<strong>of</strong> normal bladder epithelial cells at subnanomolar concentrations, leading to the<br />
hypothesis that APF could possess anticancer properties. APF was found to<br />
markedly inhibit growth <strong>of</strong> both T24 bladder carcinoma and kidney cancer cell<br />
lines. Subsequent structural and SAR studies revealed that APF (as isolated<br />
from IC/PBS patients) is a nine-residue peptide containing sialyl-TF antigen α-Olinked<br />
to the N-terminal threonine residue, Neu5Acα2-3Galβ1-3GalNAcα-O-<br />
TVPAAVVVA. Neither the sialyl group nor the C-terminal alanine residue is<br />
necessary to maintain antiproliferative activity, yet other seemingly<br />
inconsequential structural changes completely abolish the activity <strong>of</strong> APF. Thus,
we have structurally characterized APF and several APF analogues in aqueous<br />
solution using 2D NMR spectroscopic techniques in an attempt to further<br />
understand the structure <strong>of</strong> APF.<br />
CARB 108<br />
Structural and quantitative analysis <strong>of</strong> disaccharides using CE with LIF<br />
detection<br />
Yuqing Chang (1) , changy5@rpi.edu, 110 8th ST, Troy NY 12180, United States ;<br />
Tatiana Laremore (1) ; Fuming Zhang (1) ; Robert J Linhardt (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic Institute, Troy NY<br />
12180, United States<br />
Capillary electrophoresis (CE) method using laser-induced fluorescence (LIF)<br />
detection can be applied to the structural and quantitative analysis <strong>of</strong> unsaturated<br />
disaccharides. The sensitivity <strong>of</strong> this technique is attributed to the high intensity<br />
<strong>of</strong> the incident light and the ability to accurately focus the light on the capillary. By<br />
labeling the disaccharide reducing groups by proper fluorescent tags, such as<br />
fluorophore 2-aminoacridone (AMAC), and 4,4-difluoro-5,7-dimethyl-4-bora-<br />
3a,4a-diaza-s-indacene-3-propionic acid (BODIPY), hydrazide, the fluorotagged<br />
products can be separated by CE with appropriate electrophoretic conditions.<br />
The labeling process is simple and the conditions are mild. Using LIF as<br />
detection method at 488nm, the sensitivity could be largely improved compared<br />
to that was detected by UV detector at 255nm. With this strategy, it is possible to<br />
realize high sensitivity in the detection <strong>of</strong> Δ-disaccharides in trace amount.<br />
CARB 108<br />
Structural and quantitative analysis <strong>of</strong> disaccharides using CE with LIF<br />
detection<br />
Yuqing Chang (1) , changy5@rpi.edu, 110 8th ST, Troy NY 12180, United States ;<br />
Tatiana Laremore (1) ; Fuming Zhang (1) ; Robert J Linhardt (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic Institute, Troy NY<br />
12180, United States<br />
Capillary electrophoresis (CE) method using laser-induced fluorescence (LIF)<br />
detection can be applied to the structural and quantitative analysis <strong>of</strong> unsaturated<br />
disaccharides. The sensitivity <strong>of</strong> this technique is attributed to the high intensity<br />
<strong>of</strong> the incident light and the ability to accurately focus the light on the capillary. By<br />
labeling the disaccharide reducing groups by proper fluorescent tags, such as<br />
fluorophore 2-aminoacridone (AMAC), and 4,4-difluoro-5,7-dimethyl-4-bora-<br />
3a,4a-diaza-s-indacene-3-propionic acid (BODIPY), hydrazide, the fluorotagged<br />
products can be separated by CE with appropriate electrophoretic conditions.<br />
The labeling process is simple and the conditions are mild. Using LIF as
detection method at 488nm, the sensitivity could be largely improved compared<br />
to that was detected by UV detector at 255nm. With this strategy, it is possible to<br />
realize high sensitivity in the detection <strong>of</strong> Δ-disaccharides in trace amount.<br />
CARB 109<br />
Development <strong>of</strong> a novel cancer vaccine based on multivalent presentation<br />
<strong>of</strong> tumor-associated carbohydrate antigens on gold nanoparticle scaffolds<br />
Raymond P Brinas (1) , brinasr@mail.nih.gov, 376 Boyles St., Frederick MD<br />
21702, United States ; Andreas Sundgren (1) ; Micah Maetani (1) ; Omar<br />
Abbudayyeh (1) ; Howard A Young (2) ; Michael Sanford (2) ; Joseph J Barchi (1) . (1)<br />
<strong>Chemical</strong> Biology Lab, National Cancer Institute, NIH, Frederick MD 21702,<br />
United States (2) Laboratory <strong>of</strong> Experimental Biology, National Cancer Institute,<br />
NIH, Frederick MD 21702, United States<br />
We have designed and synthesized vaccine constructs based on gold<br />
nanoparticles (Au NPs) bearing multiple copies <strong>of</strong> a tumor-associated<br />
carbohydrate, Thomsen-Friedenreich (TF) disaccharide, which was O-linked to<br />
either serine or threonine residues <strong>of</strong> a specific, thiol functionalized 16-mer<br />
peptide repeating unit from a specific tumor-associated mucin (MUC4). The Au<br />
NPs were furtherfunctionalized with a segment from a form <strong>of</strong> the complementderived<br />
protein, C3d, and with the linker that was used to attach the peptides to<br />
gold. Initial results on the pro-inflammatory response <strong>of</strong> pre-stimulated mouse<br />
bone marrow-derived macrophages when treated with the vaccine constructs<br />
indicated particle-specific modulation <strong>of</strong> particular cytokine release. Guided by<br />
these results, we attempted to optimize responses by exploring differences<br />
based on particle size and coating ligand. We will present the details <strong>of</strong> Au NP<br />
synthesis, as well as optimization <strong>of</strong> our vaccine design based on the<br />
immunological assays.<br />
CARB 110<br />
Carbon nanotubes and chitosan as possible scaffolds for bone tissue<br />
regeneration<br />
Julia Stone (1) , jmstone_81@yahoo.com, PO Box 519 MS 2008, Prairie View TX<br />
77446, United States ; Yetunde Olusanya (2) ; Whitney Jones (2) ; Pasakorn<br />
Traisawatwong (1) ; Melisa Stewart (1) ; Cordella Kelly-Brown (1) ; Laura Carson (1) ;<br />
Aderemi Oki (3) ; E. Gloria C. Regisford (2) . (1) Cooperative Agricultural Research<br />
Center, Prairie View A&M University, Prairie View TX 77446, United States (2)<br />
Department <strong>of</strong> Biology, Prairie View A&M University, Prairie View TX 77446,<br />
United States (3) Department <strong>of</strong> <strong>Chemistry</strong>, Prairie View A&M University, Prairie<br />
View TX 77446, United States
Carbon nanotubes (CNTs) and chitosan exhibit characteristics necessary for<br />
successful bone tissue regeneration. An in vitro model for bone cell growth with<br />
CNTs and Chitosan added to cell culture medium <strong>of</strong> human fetal osteoblastic<br />
cells at concentrations <strong>of</strong> 0, 100, 250, 500, and 1000 ng/ml for 24 and 48 hours.<br />
SEM images exhibited no marked changes on CNTs or CTS morphology and<br />
composition. Cell proliferation was significantly higher in cells grown in the<br />
presence <strong>of</strong> 500 ng/mL <strong>of</strong> CNT, compared to controls. Western blot analysis<br />
revealed that OPN expression did not exceed that <strong>of</strong> control cells, although cells<br />
treated with 500 ng/ml CNTs showed the greatest expression. Cells treated with<br />
CTS had a dose dependent increase in OPN expression, with the highest<br />
expression at 1000 ng/ml. These data indicate that CNTs and CTS may be <strong>of</strong><br />
great potential for therapeutic use in bone regeneration.<br />
CARB 110<br />
Carbon nanotubes and chitosan as possible scaffolds for bone tissue<br />
regeneration<br />
Julia Stone (1) , jmstone_81@yahoo.com, PO Box 519 MS 2008, Prairie View TX<br />
77446, United States ; Yetunde Olusanya (2) ; Whitney Jones (2) ; Pasakorn<br />
Traisawatwong (1) ; Melisa Stewart (1) ; Cordella Kelly-Brown (1) ; Laura Carson (1) ;<br />
Aderemi Oki (3) ; E. Gloria C. Regisford (2) . (1) Cooperative Agricultural Research<br />
Center, Prairie View A&M University, Prairie View TX 77446, United States (2)<br />
Department <strong>of</strong> Biology, Prairie View A&M University, Prairie View TX 77446,<br />
United States (3) Department <strong>of</strong> <strong>Chemistry</strong>, Prairie View A&M University, Prairie<br />
View TX 77446, United States<br />
Carbon nanotubes (CNTs) and chitosan exhibit characteristics necessary for<br />
successful bone tissue regeneration. An in vitro model for bone cell growth with<br />
CNTs and Chitosan added to cell culture medium <strong>of</strong> human fetal osteoblastic<br />
cells at concentrations <strong>of</strong> 0, 100, 250, 500, and 1000 ng/ml for 24 and 48 hours.<br />
SEM images exhibited no marked changes on CNTs or CTS morphology and<br />
composition. Cell proliferation was significantly higher in cells grown in the<br />
presence <strong>of</strong> 500 ng/mL <strong>of</strong> CNT, compared to controls. Western blot analysis<br />
revealed that OPN expression did not exceed that <strong>of</strong> control cells, although cells<br />
treated with 500 ng/ml CNTs showed the greatest expression. Cells treated with<br />
CTS had a dose dependent increase in OPN expression, with the highest<br />
expression at 1000 ng/ml. These data indicate that CNTs and CTS may be <strong>of</strong><br />
great potential for therapeutic use in bone regeneration.<br />
CARB 111<br />
Biosynthesis <strong>of</strong> heparin by metabolic engineering <strong>of</strong> Chinese hamster<br />
ovary cells
Leyla Gasimli (1) , gasiml@rpi.edu, 12 Marshall Str, 1FL Rear apt, Troy NY<br />
12180, United States ; Jongyoun Baik (2) ; Susan Sharfstein (2) ; Robert J.<br />
Linhardt (1)(3)(4) . (1) Department <strong>of</strong> Biology, Rensselaer Polytechnic Institute, Troy<br />
NY 12180, United States (2) College <strong>of</strong> Nanoscale Science and Engineering,<br />
State University <strong>of</strong> New York at Albany, Albany NY 12203, United States (3)<br />
Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic<br />
Institute, Troy NY 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong> and<br />
Biological Engineering, Rensselaer Polytechnic Institute, Troy NY 12180, United<br />
States<br />
Heparin(HP), a highly sulfated polysaccharide, is a widely used anticoagulant<br />
with direct healthcare applications in the pharmaceutical market. The primary<br />
sources <strong>of</strong> heparin on the market are <strong>of</strong> animal origin. These supplies inherently<br />
involve risk <strong>of</strong> contamination in addition to uncontrollable qualitative differences<br />
among batches. Chinese hamster ovary(CHO) cells have been used as<br />
producers <strong>of</strong> functional proteins on an industrial scale. They naturally produce<br />
heparan sulfate(HS), which shares the biosynthesis pathway with HP. Therefore<br />
CHO cells are good candidates to produce clean and consistent HP on a large<br />
scale. Preliminary results show that genes coding for C5Epimerase, 2-Osulfotransferase,<br />
and 6-O- sulfotransferases are expressed in CHO cells,<br />
whereas genes coding for N-deacetylase/N-sulfotransferase-2(NDST2), and 3-Osulfotransferases(3OST)<br />
are not. NDST2 has been already transfected into cells.<br />
3OSTs will also be transfected and overexpressed in cells with the goal that HP<br />
will be synthesized on the core protein <strong>of</strong> HS, greatly simplifying large scale<br />
purification.<br />
CARB 111<br />
Biosynthesis <strong>of</strong> heparin by metabolic engineering <strong>of</strong> Chinese hamster<br />
ovary cells<br />
Leyla Gasimli (1) , gasiml@rpi.edu, 12 Marshall Str, 1FL Rear apt, Troy NY<br />
12180, United States ; Jongyoun Baik (2) ; Susan Sharfstein (2) ; Robert J.<br />
Linhardt (1)(3)(4) . (1) Department <strong>of</strong> Biology, Rensselaer Polytechnic Institute, Troy<br />
NY 12180, United States (2) College <strong>of</strong> Nanoscale Science and Engineering,<br />
State University <strong>of</strong> New York at Albany, Albany NY 12203, United States (3)<br />
Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong> Biology, Rensselaer Polytechnic<br />
Institute, Troy NY 12180, United States (4) Department <strong>of</strong> <strong>Chemical</strong> and<br />
Biological Engineering, Rensselaer Polytechnic Institute, Troy NY 12180, United<br />
States<br />
Heparin(HP), a highly sulfated polysaccharide, is a widely used anticoagulant<br />
with direct healthcare applications in the pharmaceutical market. The primary<br />
sources <strong>of</strong> heparin on the market are <strong>of</strong> animal origin. These supplies inherently<br />
involve risk <strong>of</strong> contamination in addition to uncontrollable qualitative differences<br />
among batches. Chinese hamster ovary(CHO) cells have been used as
producers <strong>of</strong> functional proteins on an industrial scale. They naturally produce<br />
heparan sulfate(HS), which shares the biosynthesis pathway with HP. Therefore<br />
CHO cells are good candidates to produce clean and consistent HP on a large<br />
scale. Preliminary results show that genes coding for C5Epimerase, 2-Osulfotransferase,<br />
and 6-O- sulfotransferases are expressed in CHO cells,<br />
whereas genes coding for N-deacetylase/N-sulfotransferase-2(NDST2), and 3-Osulfotransferases(3OST)<br />
are not. NDST2 has been already transfected into cells.<br />
3OSTs will also be transfected and overexpressed in cells with the goal that HP<br />
will be synthesized on the core protein <strong>of</strong> HS, greatly simplifying large scale<br />
purification.<br />
CARB 112<br />
Purification strategies for separation <strong>of</strong> capsular polysaccharide from<br />
fermentation broth<br />
Ujjwal Bhaskar (1) , bhasku@rpi.edu, 4117, CBIS, 110 8th Street, Troy New York<br />
12180, United States ; Zhenyu Wang (2) ; Fuming Zhang (2) ; Jonathan S.<br />
Dordick (1)(2) ; Robert J. Linhardt (1)(2) . (1) Department <strong>of</strong> <strong>Chemical</strong> and Biological<br />
Engineering, Rensselaer Polytechnic Institute, Troy New York 12180, United<br />
States (2) Center for Biotechnology and Interdisciplinary Studies, Rensselaer<br />
Polytechnic Institute, Troy New York 12180, United States<br />
In the recent past, contamination problems have arisen with heparin derived from<br />
porcine tissues. One solution to this issue is the synthesis <strong>of</strong> a bioengineered<br />
version <strong>of</strong> heparin, which could enable large-scale heparin production. Capsular<br />
polysaccharide derived from K5 strain <strong>of</strong> Escherichia coli provides a<br />
polysaccharide precursor for the development <strong>of</strong> bioengineered heparin. This<br />
study deals with the various purification strategies for the separation <strong>of</strong> the<br />
capsular polysaccharide, heparosan, from the fermentation broth <strong>of</strong> E. coli K5<br />
culture. The purification makes use <strong>of</strong> the surface charge present on the<br />
polysaccharides based on the manipulation <strong>of</strong> solution pH. Separation methods<br />
including ion-exchange chromatography and affinity precipitation are explored in<br />
heparosan purification. Affinity precipitation-based separation systems, in place<br />
<strong>of</strong> ion exchange columns, can provide an efficient, lower cost separation.<br />
Naturally derived polycations are investigated as a possible mediator <strong>of</strong> affinitybased<br />
separation for heparosan.<br />
CARB 113<br />
Design, synthesis, and characterization <strong>of</strong> sulfated N-aryl aminoglycosides<br />
Amanda M. Fenner (1) , amanda-fenner@uiowa.edu, 115 S. Grand Ave., S311<br />
PHAR, Iowa City IA 52242, United States ; Robert J. Kerns (1) . (1) <strong>Division</strong> <strong>of</strong><br />
Medicinal and Natural Products <strong>Chemistry</strong>, The University <strong>of</strong> Iowa, Iowa City IA<br />
52242, United States
Hundreds <strong>of</strong> eukaryotic and prokaryotic proteins bind to cell surface<br />
glycosaminoglycans (GAGs) to promote biological activities. Development <strong>of</strong><br />
compounds to block GAG-protein interactions has primarily focused on<br />
optimizing the degree and orientation <strong>of</strong> anionic substituents on a scaffold, to<br />
mimic GAG structure, but their utility is diminished by non-specific interactions<br />
with many proteins. To overcome these limitations, our lab demonstrated that<br />
replacing N-sulfo groups on heparin with non-anionic N-arylacyl groups<br />
increased affinity and selectivity for binding different heparin-binding proteins.<br />
Here we report the preparation and characterization <strong>of</strong> N-aryl substituted tri- and<br />
tetrasaccharides followed by sulfonation <strong>of</strong> the hydroxyl groups. An LC-MS<br />
method to analyze components <strong>of</strong> the resulting mixtures was developed using<br />
ion-pair-RP-HPLC to separate oligosaccharides based on degree and position <strong>of</strong><br />
sulfates, and ESI-MS to characterize individual molecules. Screening for<br />
biological activity indicates the aromatic substituent, core saccharide, and degree<br />
<strong>of</strong> sulfation are all important for differentiation <strong>of</strong> function.<br />
CARB 113<br />
Design, synthesis, and characterization <strong>of</strong> sulfated N-aryl aminoglycosides<br />
Amanda M. Fenner (1) , amanda-fenner@uiowa.edu, 115 S. Grand Ave., S311<br />
PHAR, Iowa City IA 52242, United States ; Robert J. Kerns (1) . (1) <strong>Division</strong> <strong>of</strong><br />
Medicinal and Natural Products <strong>Chemistry</strong>, The University <strong>of</strong> Iowa, Iowa City IA<br />
52242, United States<br />
Hundreds <strong>of</strong> eukaryotic and prokaryotic proteins bind to cell surface<br />
glycosaminoglycans (GAGs) to promote biological activities. Development <strong>of</strong><br />
compounds to block GAG-protein interactions has primarily focused on<br />
optimizing the degree and orientation <strong>of</strong> anionic substituents on a scaffold, to<br />
mimic GAG structure, but their utility is diminished by non-specific interactions<br />
with many proteins. To overcome these limitations, our lab demonstrated that<br />
replacing N-sulfo groups on heparin with non-anionic N-arylacyl groups<br />
increased affinity and selectivity for binding different heparin-binding proteins.<br />
Here we report the preparation and characterization <strong>of</strong> N-aryl substituted tri- and<br />
tetrasaccharides followed by sulfonation <strong>of</strong> the hydroxyl groups. An LC-MS<br />
method to analyze components <strong>of</strong> the resulting mixtures was developed using<br />
ion-pair-RP-HPLC to separate oligosaccharides based on degree and position <strong>of</strong><br />
sulfates, and ESI-MS to characterize individual molecules. Screening for<br />
biological activity indicates the aromatic substituent, core saccharide, and degree<br />
<strong>of</strong> sulfation are all important for differentiation <strong>of</strong> function.<br />
CARB 114<br />
Photo-click immobilization <strong>of</strong> carbohydrates on polymeric surfaces: An<br />
effortless method to functionalize surfaces for biomolecular recognition<br />
studies
Oscar Norberg (1) , oscar3@kth.se, Teknikringen 30, Stockholm 100 44, Sweden<br />
; Lingquan Deng (1) ; Mingdi Yan (2) ; Ol<strong>of</strong> Ramström (1) . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, KTH - Royal Institute <strong>of</strong> Technology, Stockholm 100 44, Sweden (2)<br />
Department <strong>of</strong> <strong>Chemistry</strong>, Portland State University, Portland, United States<br />
<strong>Carbohydrate</strong>s play highly diverse roles in biological systems. To efficiently study<br />
the interactions between carbohydrates and proteins, efforts have been made in<br />
applying microarray technology to carbohydrates. However, there are numerous<br />
obstacles to overcome concerning the manufacture <strong>of</strong> carbohydrate microarrays,<br />
where robust and versatile formats are needed.<br />
In the present study a method has been developed for efficient carbohydrate<br />
attachment to different polymeric surfaces. A specific bifunctional linker was<br />
designed for the immobilization method, efficiently connecting the polymeric<br />
substrate to the carbohydrates. The method utilizes efficient photocoupling <strong>of</strong><br />
stabilized perfluorophenyl azides (PFPAs) and highly chemoselective Copper<br />
catalyzed Azide- Alkyne Cycloaddition.<br />
The method enables a rapid and convenient protocol to the general attachment<br />
<strong>of</strong> azide-functionalized structures and was used to fabricate a range <strong>of</strong> surfaces<br />
presenting β-D-galactosides, α-D-mannosides and N-acetyl-β-D-glucosamines<br />
on different polymeric surfaces. The surfaces were evaluated in real-time studies<br />
using a QCM flow-through system with a series <strong>of</strong> different lectins.<br />
CARB 115<br />
On-line micr<strong>of</strong>low high-performance liquid chromatography with nanoelectrospray<br />
ionization mass spectrometry for heparan sulfate<br />
disaccharide analysis<br />
Bo Yang (1) , yangb4@rpi.edu; Kemal Solakyildirim (1) ; Jeffrey G. Martin (1) ; Tatiana<br />
Laremore (1) ; Robert J. Linhardt (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong><br />
Biology, Rensselaer Polytechnic Institute, Troy New York 12180, United States
The heparan sulfate and heparin, as an important kind <strong>of</strong> glycosaminoglycan<br />
(GAG), is composed acidic and linear chains <strong>of</strong> repeating multiple disaccharide<br />
units. They have crucial biological functions by binding to different growth factors,<br />
enzymes, morphogens, cell adhesion molecules, and cytokines. The<br />
characterization <strong>of</strong> HS/heparin with various sequences and structures is essential<br />
for elucidating the functions corresponding to these GAGs. Due to their structural<br />
complexity and heterogeneity, the structural analysis <strong>of</strong> these GAGs is still a<br />
long-standing challenge for analysts. In this work, a highly sensitive and fast<br />
method by ion pairing reversed-phase micr<strong>of</strong>low high-performance liquid<br />
chromatography (IPRP-Mf-HPLC) with nano-electrospray ionization mass<br />
spectrometry was applied for separation and identification <strong>of</strong> HS/ heparin<br />
disaccharides. The new approach allows us to detect approximately 0.1 ng <strong>of</strong> per<br />
disaccharide.<br />
CARB 115<br />
On-line micr<strong>of</strong>low high-performance liquid chromatography with nanoelectrospray<br />
ionization mass spectrometry for heparan sulfate<br />
disaccharide analysis<br />
Bo Yang (1) , yangb4@rpi.edu; Kemal Solakyildirim (1) ; Jeffrey G. Martin (1) ; Tatiana<br />
Laremore (1) ; Robert J. Linhardt (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and <strong>Chemical</strong><br />
Biology, Rensselaer Polytechnic Institute, Troy New York 12180, United States<br />
The heparan sulfate and heparin, as an important kind <strong>of</strong> glycosaminoglycan<br />
(GAG), is composed acidic and linear chains <strong>of</strong> repeating multiple disaccharide<br />
units. They have crucial biological functions by binding to different growth factors,<br />
enzymes, morphogens, cell adhesion molecules, and cytokines. The<br />
characterization <strong>of</strong> HS/heparin with various sequences and structures is essential<br />
for elucidating the functions corresponding to these GAGs. Due to their structural<br />
complexity and heterogeneity, the structural analysis <strong>of</strong> these GAGs is still a<br />
long-standing challenge for analysts. In this work, a highly sensitive and fast<br />
method by ion pairing reversed-phase micr<strong>of</strong>low high-performance liquid<br />
chromatography (IPRP-Mf-HPLC) with nano-electrospray ionization mass<br />
spectrometry was applied for separation and identification <strong>of</strong> HS/ heparin<br />
disaccharides. The new approach allows us to detect approximately 0.1 ng <strong>of</strong> per<br />
disaccharide.<br />
CARB 116<br />
De novo synthesis <strong>of</strong> a 2-acetamido-4-amino-2,4,6-trideoxy-D-galactose<br />
(AAT) building block for the preparation <strong>of</strong> the zwitterionic polysaccharide<br />
A1 (PS A1) repeating subunit <strong>of</strong> Bacteroides fragilis<br />
Rajan Pragani (1)(2) , rajan.pragani@mpikg.mpg.de, Am Muehlenberg 1, Potsdam<br />
Brandenburg 14476, Germany ; Peter H Seeberger (1)(2) . (1) Department <strong>of</strong>
Biomolecular Systems, Max-Planck-Institute <strong>of</strong> Colloids and Interfaces, Potsdam<br />
14476, Germany (2) Department <strong>of</strong> <strong>Chemistry</strong> and Biochemistry, Freie<br />
Universitaet Berlin, Potsdam 14476, Germany<br />
Zwitterionic polysaccharide A1 (PS A1) found in Bacteroides fragilis is the<br />
putative cause <strong>of</strong> postoperative intra-abdominal abscesses in humans and has<br />
been shown to activate a MHCII mediated T-cell response in the absence <strong>of</strong><br />
proteins. Total synthesis <strong>of</strong> defined homogeneous PS A1 fragments would<br />
enable a more in-depth understanding <strong>of</strong> this important immunostimulant. Herein,<br />
we present our progress toward the synthesis <strong>of</strong> the PS A1 repeating subunit.<br />
Detailed accounts <strong>of</strong> the de novo synthesis <strong>of</strong> the 2-acetamido-4-amino-2,4,6trideoxy-D-galactose<br />
(AAT) monosaccharide, and its use toward the completion<br />
<strong>of</strong> the PS A1 repeating subunit will be reported.<br />
CARB 117<br />
Automated solution-phase synthesis <strong>of</strong> alpha-galactosides<br />
Rajarshi Roychoudhury (1) , rajarshi@iastate.edu, 2612 Gilman, Ames IA 50011,<br />
United States ; Nicola L. B. Pohl (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong> and The Plant<br />
Sciences Institute, Iowa State University, Ames IA 50011, United States<br />
Human cells do not express alpha-galactoside (alpha-Gal) epitopes. However,<br />
constant exposure to this epitope leads to humans producing high levels <strong>of</strong><br />
antibodies against this carbohydrate motif. These antibodies can be used to<br />
target immune responses against other compounds that are conjugated to alpha-<br />
Gal. Unfortunately, the synthesis <strong>of</strong> this motif is particularly challenging as<br />
anchimeric assistance cannot be used to set the correct anomeric configuration.<br />
Here we present a systematic study <strong>of</strong> several activated galactose building<br />
blocks to ascertain the influence <strong>of</strong> conformational effects in determining the<br />
alpha/beta selectivity in glycosylation reactions. We also demonstrate the<br />
incorporation <strong>of</strong> the most selective building block in the development <strong>of</strong> the first<br />
automated solution-phase synthesis protocol for the incorporation <strong>of</strong> terminal<br />
alpha-galactosides.<br />
CARB 118<br />
Study <strong>of</strong> the protecting group participation and stereoselectivity <strong>of</strong> 2-azido-<br />
2-deoxygalactopyranosyl donors<br />
Zhitao Li (1) , zli@binghamton.edu, Binghamton University, Binghamton NY<br />
13902, United States . (1) Department <strong>of</strong> <strong>Chemistry</strong>, Binghamton University,<br />
Binghamton NY 13902, United States<br />
Protecting groups have pr<strong>of</strong>ound effects on the stereoselectivity <strong>of</strong> glycosylation<br />
reactions. These effects include neighboring group participation, electronic effect
and steric effect. Nonneighboring group participation was recently reported to be<br />
responsible for stereoselectivity in some glycosylation reactions. Our<br />
computational model study indicated that the nonneighboring group participation<br />
may also contribute to the stereoselectivity <strong>of</strong> glycosylation <strong>of</strong> 2-azido-2deoxygalactosyl<br />
(GalN3) donors. A series <strong>of</strong> GalN3 donors with different<br />
protecting groups were synthesized and tested in glycosylation reactions. The<br />
relationship between the protecting pattern and the stereoselectivity <strong>of</strong> the<br />
reaction is consistent with the predictions based on our computational model.<br />
This result suggests that our theoretical model is effective and may be applied to<br />
develop more stereoselective donors.<br />
CARB 119<br />
Reactions <strong>of</strong> glycals with ortho-substituted benzanilines<br />
Cecilia H Marzabadi (1) , cecilia.marzabadi@shu.edu, 400 South Orange Ave,<br />
South Orange NJ 07079, United States ; Katherine Kochalski (1) . (1) <strong>Chemistry</strong><br />
and Biochemistry, Seton Hall University, South Orange NJ 07079, United States<br />
Previously we reported on the successful reactions <strong>of</strong> glycals with benzanilines to<br />
form carbohydrate-based quinolines and tetrahydroquinolines. We were<br />
interested to see the outcome <strong>of</strong> these addition reactions when a nucleophilic<br />
group was substituted on the ortho-position <strong>of</strong> the aldehyde ring. In this<br />
presentation we will discuss the results <strong>of</strong> these studies and our efforts to<br />
prepare pyranobenzopyrans and other heterocycles from carbohydrate<br />
precursors. [ACS_F09_Marzabadi]<br />
CARB 120<br />
Novel glycolipids in CD1d-mediated immunity: Synthesis <strong>of</strong> new agonists<br />
for CD1d<br />
Justyna Wojno (1)(2) , jxw732@bham.ac.uk, Edgbaston, Birmingham West<br />
Midlands B15 2TT, United Kingdom ; John-Paul Jukes (3) ; Paolo Polzella (3) ;<br />
Vincenzo Cerundolo (3) ; Liam R Cox (1) ; Gurdyal S Besra (2) . (1) School <strong>of</strong><br />
<strong>Chemistry</strong>, University <strong>of</strong> Birmingham, Birmingham B15 2TT, United Kingdom (2)<br />
School <strong>of</strong> Biosciences, University <strong>of</strong> Birmingham, Birmingham B15 2TT, United<br />
Kingdom (3) Tumour Immunology Group, Weatherall Institute <strong>of</strong> Molecular<br />
Medicine, John Radcliffe Hospital, Oxford OX1 3QT, United Kingdom
The glycolipid α-galactosyl ceramide, α-GalCer, has been shown to stimulate the<br />
proliferation <strong>of</strong> murine spleen cells and activate the immune system through<br />
binding to the protein CD1d. The induction <strong>of</strong> both Th1 and Th2 cytokines by α-<br />
GalCer is likely to limit its therapeutic utility. Analogues <strong>of</strong> α-GalCer have been<br />
shown to induce iNKT cell-derived cytokines more selectively through a skewed<br />
Th1-Th2 response. The crystal structure <strong>of</strong> human CD1d in complex with α-<br />
GalCer revealed a key hydrogen bond between the N-H <strong>of</strong> the amide group and<br />
the CD1d protein but the C=O is not involved in direct binding. A range <strong>of</strong> αgalactosyl<br />
ceramide and threitol ceramide analogues have been synthesised in<br />
which the amide group in our lead has been replaced with different carbonyl<br />
functional groups. Additional modifications will also be presented along with the<br />
synthesis <strong>of</strong> labelled derivatives, which will find application in trafficking studies.<br />
CARB 121<br />
Automated synthesis <strong>of</strong> systematic di- and tri-saccharide libraries<br />
Xin Liu (1) , xinl@iastate.edu, 2606 Gilman, Ames IA 50011, United States ;<br />
Beatrice Y. M. Collet (2) ; Shu-Lun Tang (1) ; Sahana K. Nagappayya (1) ; Rajarshi<br />
Roychoudhury (1) ; Xueshu Li (1) ; Heather Edwards (1) ; Nicola L. B. Pohl (1) . (1)<br />
Department <strong>of</strong> <strong>Chemistry</strong> and The Plant Sciences Institute, Iowa State University,<br />
Ames IA 50011, United States (2) LuCella Biosciences, Inc., Ames IA 50011,<br />
United States<br />
The lack <strong>of</strong> availability <strong>of</strong> small carbohydrate libraries with systematic structural<br />
variations has limited a rigorous study <strong>of</strong> the role <strong>of</strong> stereochemical features on<br />
the biological activity <strong>of</strong> carbohydrates as well as the development <strong>of</strong> robust<br />
methods such as mass-spectrometry-based carbohydrate sequencing, NMR<br />
structural databases, and predictive computational tools for carbohydrate<br />
structures. To address these issues as well as probe the scope and limitations <strong>of</strong><br />
automated oligosaccharide synthesis, the synthesis <strong>of</strong> the first di- and trisaccharide<br />
library is reported with particular focus on the incorporation <strong>of</strong> the<br />
mass identical monomers glucose, galactose, and mannose. The large-scale<br />
preparation and process issues involved in the synthesis <strong>of</strong> monosaccharide<br />
building blocks necessary for the synthesis <strong>of</strong> the library are also discussed.<br />
CARB 122<br />
Probe <strong>of</strong> activated glycoside building block stability for automated<br />
solution-phase synthesis <strong>of</strong> carbohydrate libraries<br />
Beatrice Y. M. Collet (1) , bcollet@lucellabio.com, 1030 Roy J. Carver Co-Lab,<br />
Ames IA 50011, United States ; Xin Liu (2) ; Shu-Lun Tang (2) ; Heather Edwards (2) ;<br />
Sahana Nagappayya (2) ; Lin Liu (2) ; Nicola L. B. Pohl (2) . (1) LuCella Biosciences,<br />
Inc., Ames IA 50011, United States (2) Department <strong>of</strong> <strong>Chemistry</strong> and The Plant<br />
Sciences Institute, Iowa State University, Ames IA 50011, United States
Trichloroacetimidate-activated glycosides are commonly used as donor building<br />
blocks in the synthesis <strong>of</strong> carbohydrate-based macromolecules. However the<br />
stability <strong>of</strong> these building blocks is <strong>of</strong>ten debated and could potentially limit their<br />
use in automated synthesis protocols. In the context <strong>of</strong> the automated solutionphase<br />
synthesis <strong>of</strong> di- and tri-saccharide libraries, the preparation <strong>of</strong> a variety <strong>of</strong><br />
Schmidt trichloroacetimidate donors with systematic structural variations will be<br />
reported. The stability <strong>of</strong> these building blocks at four different storage<br />
temperatures will be presented and structure-related patterns in the degradation<br />
processes will be proposed.<br />
CARB 123<br />
Synthesis <strong>of</strong> unnatural glycosaminoglycans and evaluation <strong>of</strong> their<br />
interaction with proteins<br />
Smritilekha Bera (1) , lekha026@yahoo.com, 110, 8th St, BIOTECH, 4005, Troy<br />
NY 1280, United States . (1) Department <strong>of</strong> chemistry and <strong>Chemical</strong> Biology,<br />
Rensselaer Polytechnic Institute, Troy NY 12180, United States<br />
Glycosaminoglycans (GAGs) play a wide range <strong>of</strong> physiological and pathological<br />
events as a decisive agent for cell-cell communications, cell proliferation,<br />
angiogenesis, inflammatory processes, wound repair and healing, viral invasion<br />
as well as a growth factor receptor and enzyme inhibitors and so on. These<br />
molecules are linear, anionic and having repetitive disaccharide motifs and<br />
discriminated in between and within GAG families by structural architecture with<br />
slight variations in stereochemistry, length, and patterns <strong>of</strong> sulfation. This<br />
structural diversity creates an enormous number <strong>of</strong> protein binding motifs and<br />
also has featured daunting challenge to define the structural and functional<br />
properties <strong>of</strong> GAGs, which could be implicated in many diseases and targets <strong>of</strong><br />
therapeutic approaches.<br />
Heparin sulfate, one <strong>of</strong> the most important natural glycosaminoglycans, is used<br />
as anticoagulant drug and exploited in treating cancers and suppressing<br />
inflammatory responses and chondroitin sulfate (CS), an another essential<br />
glycosaminoglycans play vital roles in biological processes, such as neural<br />
development, viral invasion, cancer metastasis, arthritis, and spinal cord injury<br />
etc. Although, these GAGs are generally isolated from living animals, synthetic<br />
chemistry <strong>of</strong>fers a potentially more reliable and practicable route to<br />
homogeneous glycosaminoglycan leading to either exact copies <strong>of</strong> naturally<br />
occurring glycosaminoglycan or, alternatively, incorporating unnatural<br />
proteolytically stable glycosidic linkages. Thus in a word, these artificially<br />
designed glycosaminoglycans might enhance its in vivo potentiality by retaining<br />
the geometric and spatial characteristics <strong>of</strong> the native polysaccharides and<br />
exhibiting stability toward N- and O-glycosyl hydrolase activity or inhibiting these<br />
enzymes to some extent. In addition, the artificial linkage may be finely tuned as<br />
an inert functionality during synthetic transformations within the synthesis <strong>of</strong>
target molecule. Our initial target is the synthesis <strong>of</strong> unnaturally linked<br />
glycosaminoglycans <strong>of</strong> heparin and condroitin sulfate subsequently the<br />
investigation <strong>of</strong> their interaction with protein.<br />
CARB 124<br />
Grafting <strong>of</strong> cellulose esters by single electron transfer living radical<br />
polymerization Set-LrP<br />
Petr Vlcek (1) , vlcek@imc.cas.cz, Heyrovsky Sq. 2, Praha 16206, Czech Republic<br />
; Vladimir Raus (2) ; Miroslav Janata (3) ; Jaroslav Kriz (4) ; Petra Latalova (5) ; Eva<br />
Cadova (6) . (1) Department <strong>of</strong> Controlled Polymerization, Institute <strong>of</strong><br />
Macromolecular <strong>Chemistry</strong>, Praha 16206, Czech Republic (2) Department <strong>of</strong><br />
Controlled Polymerization, Institute <strong>of</strong> Macromolecular <strong>Chemistry</strong>, Czech<br />
Republic (3) Department <strong>of</strong> Controlled Polymerization, Institute <strong>of</strong><br />
Macromolecular <strong>Chemistry</strong>, Czech Republic (4) Department <strong>of</strong> structure analysis,<br />
Institute <strong>of</strong> Macromolecular <strong>Chemistry</strong>, Czech Republic (5) Department <strong>of</strong><br />
Controlled Polymerization, Institute <strong>of</strong> Macromolecular <strong>Chemistry</strong>, Czech<br />
Republic (6) Department <strong>of</strong> Controlled Polymerization, Institute <strong>of</strong><br />
Macromolecular <strong>Chemistry</strong>, Czech Republic<br />
Cellulose diacetate and acetate butyrate were functionalized with 2bromoisobutyrylbromide<br />
or dichloroacetylchloride, giving polyfunctional<br />
macroinitiators for controlled radical polymerization, having various number <strong>of</strong><br />
functional groups. Recently, they have been used for controlled grafting by<br />
ATRP. Here, the macroiniator were grafted with (meth)acrylates and their<br />
mixtures and this polymerization has been initiated with the Percec's system,<br />
composed <strong>of</strong> zero-valent copper (fine powder) and amine-type ligands in polar<br />
solvent. The grafting proceeds as a controlled proces, semilogarithmic<br />
dependence <strong>of</strong> monomer consumption on reaction time is linear. The grafts are<br />
capable <strong>of</strong> polymerization <strong>of</strong> another monomer, giving rise to block copolymertype<br />
grafts. The process is relatively fast and the formed products contain<br />
virtually negligible amount <strong>of</strong> residual catalyst components and, therefore, they<br />
do not require additional purification by repeated precipitation or column<br />
chromatography. Thus, the SET-LRP is a convenient method for controlled<br />
synthesis <strong>of</strong> graft copolymers with cellulose, and perhaps, also other backbone.<br />
CARB 125<br />
Solvation and hydrolysis <strong>of</strong> cellulose with transition metal salts<br />
Veronika Viner (1) , benjamin.g.harvey@navy.mil, 1900 North Knox Rd., Stop<br />
#6303, China Lake CA 93555, United States ; Benjamin G. Harvey (1) ,<br />
benjamin.g.harvey@navy.mil, 1900 North Knox Rd., Stop #6303, China Lake CA<br />
93555, United States ; Roxanne L Quintana (1) . (1) Resarch Department,<br />
<strong>Chemistry</strong> <strong>Division</strong>, NAVAIR-NAWCWD, China Lake CA 93555, United States
The solvation and hydrolysis <strong>of</strong> cellulose to sugar oligomers and glucose in<br />
concentrated aqueous solutions <strong>of</strong> FeCl3 and ZnCl2 were studied. The effect <strong>of</strong><br />
the concentrations <strong>of</strong> the metal salts and cellulose, reaction temperatures, and<br />
incorporation <strong>of</strong> additional acid catalysts was examined. The salts could be<br />
conveniently separated from the products by complexation with ether, allowing<br />
for almost quantitative recovery. The products were analyzed by UV-Vis<br />
spectroscopy, HPLC, and ICP-AES to determine product distributions,<br />
conversions, and the concentrations <strong>of</strong> metal impurities in the products. High salt<br />
concentrations protected the glucose from further reaction, allowing for excellent<br />
yields <strong>of</strong> glucose under moderate conditions. To address more realistic<br />
feedstocks, heterogeneous sources <strong>of</strong> cellulose, including wood and paper, were<br />
subjected to optimized hydrolysis conditions. Both feedstocks were readily<br />
solvated with moderate conversion to glucose. These results are promising for<br />
the development <strong>of</strong> an efficient system for the conversion <strong>of</strong> waste biomass to<br />
glucose.<br />
CARB 126<br />
<strong>Chemical</strong> derivatization <strong>of</strong> glucan microparticles for targeted delivery<br />
Ernesto R Soto (1) , Ernesto.Soto@umassmed.edu, 373 Plantation Street,<br />
Biotech 2, Suite 113, Worcester MA 01604, United States ; Gary R Ostr<strong>of</strong>f (1) . (1)<br />
Program in Molecular Medicine, University <strong>of</strong> Massachusetts Medical School,<br />
Worcester MA 01604, United States<br />
We have previously reported on targeted delivery <strong>of</strong> siRNA to macrophages<br />
using siRNA encapsulated β-glucan particles (GP). GP are 2-4 μm hollow and<br />
porous microspheres that facilitate uptake by cells bearing β-glucan receptors<br />
(dectin-1 and CR3) thus providing for targeted delivery to innate immune cells<br />
(macrophages, dendritic cells, and neutrophils). To extend the application <strong>of</strong> the<br />
GP delivery technology to non-pr<strong>of</strong>essional phagocytic cells, we have developed<br />
methods to chemically graft targeting ligands to the glucan particle surface to<br />
specific cell surface receptors. We will detail the methods and results <strong>of</strong><br />
chemically modifying GPs with galactose to target particles to hepatocytes via<br />
the asialoglycoprotein receptor, or with arginine–glycine–aspartic acid (RGD)containing<br />
peptides to target particles to endothelial cells and fibroblasts via the<br />
β1 integrin receptor.<br />
CARB 127<br />
Degradation mechanism and crystal structure change <strong>of</strong> cellulose during<br />
TEMPO-NaOCl-NaBr selective oxidation and its biomedical application as<br />
hemostatics<br />
Bin Sun (1) , sunbin7210@gmail.com, 2999 North Renmin Road,Songjiang, Room<br />
C402,Buld 5, Shanghai Shanghai 201620, China ; Wenxiu Han (2) ; Wei Wang (2) ;
Jinhong Ma (1) ; Chunju Gu (1) ; Meifang Zhu (1) ; Borun Liang (1) . (1) State Key Lab for<br />
Modification <strong>of</strong> <strong>Chemical</strong> Fibers and Polymer Materials, College <strong>of</strong> Material<br />
Science and Engineering, Donghua University, Shanghai 201620, China (2) Key<br />
Laboratory <strong>of</strong> Science and Technology <strong>of</strong> Eco-textile, Ministry <strong>of</strong> Education,<br />
College <strong>of</strong> <strong>Chemical</strong> Science and Engineering, Donghua University, Shanghai<br />
201620, China<br />
The functional modification <strong>of</strong> cellulose and its application in the field <strong>of</strong><br />
biomedical materials has been considered as one <strong>of</strong> the most important research<br />
directions, which attracted much attention in recent years. In this contribution,the<br />
degradation extent and its mechanism <strong>of</strong> the cellulose during TEMPO-NaOCl-<br />
NaBr selective oxidation process have been explored. The crystal structure and<br />
absorbable hemostatic properties <strong>of</strong> the celluronic acid have been characterized<br />
in detail. It is suggested that the significant degradation at the initial stage is<br />
certainly associated with the fringed-micelle structure that rayon fiber has and the<br />
β-elimination induced by the formation <strong>of</strong> the aldehyde intermediate in the<br />
alkaline solution. The fully oxidized celluronic acid sodium salt exhibits<br />
amorphous structure in character whereas the celluronic acid obtained after<br />
treating with acid exhibits clear crystal structure in character, which is quite<br />
similar to the crystal modification <strong>of</strong> cellulose .<br />
The change <strong>of</strong> the hydrogen bond structure owing to the introduction <strong>of</strong> the<br />
carboxyl group at C6 position was discussed.
The results upon preliminary animal test indicate that the oxidized cellulose<br />
products have good hemostatic and absorbable properties which has rapid<br />
hemostasis in 2 minutes, and is basically absorbed in the body <strong>of</strong> rabbits within 6<br />
days.<br />
CARB 128<br />
Temperature-dependent chain-collapse behavior <strong>of</strong> cellulose ethers in<br />
dilute solution<br />
Hongwei Shen (1) , hshen@dow.com, 2301 N Brazosport Blvd., B-1220/Rm101,<br />
Freeport TX 77541, United States ; Robert Sammler (1) ; David Redwine (1) ; David<br />
Meunier (1) ; Meinolf Brackhagen (1) . (1) The Dow <strong>Chemical</strong> Company, Freeport TX<br />
77541, United States<br />
Chain behavior <strong>of</strong> cellulose ethers (polysaccharides) in dilute aqueous solution<br />
was studied using light scattering techniques. Radius <strong>of</strong> gyration (Rg) and<br />
hydrodynamic radius (Rh) were determined for various types <strong>of</strong> cellulose ethers<br />
over a range <strong>of</strong> concentrations and temperatures. A critical association<br />
concentration (CAC) well below its chain overlap concentration (C*) was found.<br />
This suggests polymer chain is in an associated state even below C*. Polymer<br />
chains at a concentration between CAC and C* are random coils at room<br />
temperature but form aggregates as temperature increases above a critical<br />
value. Below CAC, at a few ppm, Rg and Rh can decrease significantly with<br />
increasing temperature above a critical value and the polymer chain changes<br />
from random coils to constrained coils to spherical shape. This is the first<br />
experimental evidence on chain collapse for cellulose ethers. The degree <strong>of</strong><br />
chain collapse is discussed in term <strong>of</strong> chain stiffness, the persistence length.<br />
CARB 129<br />
Improving dyeability <strong>of</strong> nylon wiith nano-chitosan<br />
Yau Shan Szeto (1) , tcsetoys@polyu.edu.hk, Hung Hom, Kowloon, Hong Kong<br />
Special Administrative Region <strong>of</strong> China; Ho Yan Wong (1) ; Li Wei Liu (1) ; Hoi Ying<br />
Lui (1) . (1) Institute <strong>of</strong> Textiles $ Clothing, The Hong Kong Polytechnic University,<br />
Hung Hom, Hong Kong Special Administrative Region <strong>of</strong> China<br />
The dyeability <strong>of</strong> nylon treated with chitosan nano-particles was studied. Nanoparticles<br />
<strong>of</strong> chitosan were applied onto the surface <strong>of</strong> nylon fabric using a paddry-cure<br />
method. The chitosan-treated and the untreated nylon fabrics were dyed<br />
using acid dyes with different numbers <strong>of</strong> sulphonic acid groups at 1%, 2% and<br />
4% depths respectively. It was found that the rate <strong>of</strong> dyeing and the final<br />
exhaustion <strong>of</strong> the chitosan-treated nylon fabric were increased. The effect <strong>of</strong> dye<br />
structures on exhaustion was also studied. Regarding the physical property <strong>of</strong>
the fabric, the coefficient <strong>of</strong> friction <strong>of</strong> the chitosan-treated fabric was similar to<br />
that <strong>of</strong> the control. The colour fastness to water (ISO105-E01:1994) <strong>of</strong> the dyed<br />
fabrics was also assessed. It was shown that treatment with chitosan did not lead<br />
to deterioration <strong>of</strong> the colour fastness to water <strong>of</strong> nylon. In summary, the study<br />
indicated that the depth <strong>of</strong> dyeing <strong>of</strong> nylon could be increased with the chitosan<br />
treatment.<br />
CARB 130<br />
Modeling <strong>of</strong> Congo red adsorption on a surface <strong>of</strong> crystalline cellulose<br />
using molecular dynamics<br />
Miroslaw Wyszomirski (1) , mwyszomirski@ath.bielsko.pl, Willowa 2, Bielsko-<br />
Biala, Poland ; Karim Mazeau (2) . (1) Faculty <strong>of</strong> Environmental and Materials<br />
Sciences, University <strong>of</strong> Bielsko-Biala, Bielsko-Biala 43-309, Poland (2) Centre de<br />
Recherches sur les Macromolécules Végétales CNRS, Grenoble 38041, France<br />
Congo red is an azo-dye which interacts with cellulose surface. Understanding<br />
these interactions might help understanding <strong>of</strong> adsorption <strong>of</strong> cellulose binding<br />
modules <strong>of</strong> cellulases, because they share the same hydrophobic mechanisms,<br />
with implications for cellulose hydrolysis. In our efforts to identify key variables<br />
we modeled interactions between Congo red and cellulose micr<strong>of</strong>ibril in explicit<br />
aqueous environment using molecular dynamics (Accelrys Materials Studio ). We<br />
found structural requirements for these interactions to understand ruling<br />
mechanisms and to establish likely binding sites on cellulose surfaces.<br />
CARB 131<br />
Evaluation <strong>of</strong> glycopolymers and glyconanoparticles for biosensing and<br />
gene delivery applications
Ravin Narain (1) , narain@ualberta.ca, 116 St and 85 Ave, Edmonton AB, Canada<br />
; Marya Ahmed (1) . (1) <strong>Chemical</strong> and Materials Engineering, University <strong>of</strong> Alberta,<br />
Edmonton AB T6G 2G6, Canada<br />
Synthetic carbohydrate based nano-structured materials are receiving an<br />
enormous attention in recent years since those materials have been shown to<br />
mimic natural polysaccharides, glycopeptides and glycoproteins in biological<br />
systems. Peptides and proteins with carbohydrate moieties attached to their<br />
backbone have many desirable biological properties, but their chemical synthesis<br />
is a technical challenge. Therefore, designing carbohydrate based materials that<br />
can perform similar functions as glycoproteins or can recognize glycoproteins is a<br />
viable approach for better understanding <strong>of</strong> the cellular recognition processes<br />
such as uptake, cell growth regulation, differentiation, adhesion, cellular<br />
trafficking, inflammation by bacteria and viruses and the immune response.<br />
<strong>Carbohydrate</strong> based materials for biomedical applications have other benefits<br />
such as improving the solubility <strong>of</strong> the materials in physiological conditions,<br />
imparting biocompatibility in the materials (especially for drug and gene delivery<br />
applications), and enhancing specificity (e.g. glycopolymers with galactose<br />
residues have higher recognition towards hepatocytes). Recently, we have made<br />
significant advances in the design <strong>of</strong> biologically relevant glycopolymers and<br />
glyconanoparticles. For instance, we have studied the specific interactions <strong>of</strong><br />
carbohydrate residues with specific types <strong>of</strong> lectins. We have also studied the<br />
biocompatibility, the biomolecular recognition processes, cellular uptake and<br />
transfection efficiencies <strong>of</strong> the glyconanoparticles. In this talk, I will summarize on<br />
the preparation and biological applications <strong>of</strong> well-defined glycopolymers and<br />
monodisperse glyconanoparticles.<br />
CARB 132<br />
Application <strong>of</strong> data mining techniques to differentiate glucose-containing<br />
disaccharide ions fragmented via infrared-multiple photon dissociation<br />
using tunable lasers and Fourier transform ion cyclotron resonance mass<br />
spectrometry<br />
Mohammad U. Ehsan (1) , mehsan@earthlink.net, PO Box 117200, Gainesville<br />
FL 32611, United States ; Sarah Stefan (1) ; Alexander Aksenov (1) ; Vladimir<br />
Boginksi (1) ; Brad Bendiak (2) ; John Eyler (1) . (1) Department <strong>of</strong> <strong>Chemistry</strong>,<br />
University <strong>of</strong> Florida, Gainesville FL 32611, United States (2) Department <strong>of</strong> Cell<br />
and Developmental Biology, Denver Colorado, United States<br />
Data mining techniques were used to analyze mathematically the infrared<br />
multiple photon dissociation (IRMPD) fragmentation patterns <strong>of</strong> gas-phase<br />
lithiated disaccharide isomers irradiated with both a tunable CO2 laser and a free<br />
electron laser (FEL). The IR fragmentation patterns over the wavelength range <strong>of</strong><br />
9.2 to 10.6 µm have been shown in earlier work to give diagnostic fragmentation<br />
and showed distinct correlation with geometry <strong>of</strong> the anomeric carbon.
Application <strong>of</strong> data mining approaches for data analysis allowed for unambiguous<br />
determination <strong>of</strong> anomeric carbon configurations for each disaccharide isomer<br />
pair using fragmentation data for a single wavelength. This combination <strong>of</strong><br />
wavelength-selective IRMPD and data mining may <strong>of</strong>fer a powerful and<br />
convenient tool for structural differentiation <strong>of</strong> gas-phase carbohydrate<br />
complexes.<br />
CARB 133<br />
Influence <strong>of</strong> the structure <strong>of</strong> phosphoramidates on flame retardant<br />
properties <strong>of</strong> cellulose<br />
Victoria Salimova (1) , viktoriya.salimova@empa.ch, Lerchenfeldstrasse 5, Sankt<br />
Gallen Sankt Gallen 9014, Switzerland ; Sabyasachi Gaan (1) ; Joëlle<br />
Grützmacher (2) . (1) Department <strong>of</strong> Advanced Fibers, Empa, Materials Science<br />
and Technology, Sankt Gallen 9014, Switzerland (2) Department <strong>of</strong> Inorganic<br />
<strong>Chemistry</strong>, ETH Swiss Federal Institute <strong>of</strong> Technology, Zurich 8092, Switzerland<br />
The most effective flame retardants for cellulose contain phosphorus and<br />
nitrogen, either in the same compounds (Pyrovatex® CP, Ciba) or separate<br />
compounds (Proban® CC, Rhodia). Phosphorus in various chemical states<br />
interact with cellulose through acidic intermediates to reduce its flammability, and<br />
nitrogen is believed to enhance the action <strong>of</strong> phosphorus through the synergistic<br />
effect. Although numerous investigations have been performed on the reaction<br />
mechanisms <strong>of</strong> phosphorus-containing compounds, the exact mechanism <strong>of</strong><br />
action was not elucidated so far. It has been shown, that different classes <strong>of</strong><br />
organophosphorus compounds exhibit different flame retardant behaviour, when<br />
combined with cellulose. This research focuses on the effect <strong>of</strong> phosphoramidate<br />
structure on thermal decomposition and burning behavior <strong>of</strong> cellulose. For this<br />
study, dimethyl- (DMP), diethyl- (DEP), di-n-butyl- (DBP), di-isopropyl- (DIP) and<br />
diphenyl-phosphoramidates (DPP) were taken. The structures <strong>of</strong> these<br />
phosphoramidates differ in the nature <strong>of</strong> the ester group that, as it was shown,<br />
has a significant influence on the flame retardant properties <strong>of</strong> treated cellulose.<br />
Dimethyl-, di-n-butyl-, and di-isopropylphosphoramidates were synthesized<br />
according to the Todd-Atherton reaction. Cellulose in form <strong>of</strong> cotton fabrics and<br />
microcrystalline cellulose (Avicel © PH-101) were used in this study. Fabrics as<br />
well as microcrystalline cellulosic powder were impregnated with the<br />
phosphoramidates from ethanol solution and dried in the oven at 60ºC for 2<br />
hours. The amount <strong>of</strong> phosphorus retained on the fabrics and powder was<br />
controlled using elemental analysis.<br />
The burning behavior <strong>of</strong> phosphoramidates-treated samples was estimated using<br />
Limiting Oxygen Index Test. The highest values <strong>of</strong> LOI and burning rate were<br />
found to be for DMP-treated cellulose. Thermal behaviour was evaluated using<br />
thermogravimetric analysis (TGA) and microscale combustion calorimetry (MCC).
The results from these measurements correlated well with LOI values. DMPtreated<br />
cellulose exhibited lowest decomposition temperature, highest yield <strong>of</strong><br />
char at 600ºC and lowest heat release rates.<br />
The surface <strong>of</strong> the chars left after the LOI test was investigated using Scanning<br />
Electron Microscopy (SEM) and elemental analysis. The DMP-, DEP-, DIP- and<br />
DPP-chars were shown to have similar morphologies, whereas on the surface <strong>of</strong><br />
DBP-char some swellings were observed. Elemental analysis <strong>of</strong> fabrics before<br />
combustion and their chars after combustion revealed that during the burning<br />
process nothing else but cellulose is burned away, leaving the polymeric coating<br />
formed from phosphoramidate.<br />
Further attempts to get an insight into the mode <strong>of</strong> actions <strong>of</strong> phosphoramidates<br />
were contrived using evolved gas analysis techniques such as TGA-FTIR-MS.<br />
Here, gases evolved during the TGA were fed to the FT-IR gas cell and mass<br />
spectrometer over a pre-heated transfer line. Analysis <strong>of</strong> those gases revealed<br />
that different reactions are taking place phosphoramidates with distinctive<br />
structure react with cellulose.<br />
CARB 134<br />
High-flux thin-film nan<strong>of</strong>ibrous composite ultrafiltration membranes<br />
containing cellulose barrier layer<br />
Hongyang Ma (1) , honma@notes.cc.sunysb.edu, Stony Brook University, Stony<br />
Brook University New York 11794, United States ; Benjamin S. Hsiao (1) ,<br />
bhsiao@notes.cc.sunysb.edu, Stony Brook University, Stony Brook New York<br />
11794, United States ; Benjamin Chu (1) , bchu@notes.cc.sunysb.edu, Stony<br />
Brook University, Stony Brook New York, United States . (1) Department <strong>of</strong><br />
<strong>Chemistry</strong>, Stony Brook University, Stony Brook New York 11794, United States<br />
A novel class <strong>of</strong> thin-film nan<strong>of</strong>ibrous composite (TFNC) membrane consisting <strong>of</strong><br />
a cellulose barrier layer, a nan<strong>of</strong>ibrous mid-layer scaffold, and a melt-blown nonwoven<br />
substrate was successfully fabricated and tested as an ultrafiltration (UF)<br />
filter to separate an emulsified oil and water mixture, a model bilge water for onboard<br />
ship bilge water purification. Two ionic liquids: 1-butyl-3-methylimidazolium<br />
chloride and 1-ethyl-3-methylimidazolium acetate, were chosen as the solvent to<br />
dissolve cellulose under mild conditions. The permeation flux <strong>of</strong> the cellulosebased<br />
TFNC membrane was significantly higher (e.g. 10X) than comparable<br />
commercial UF membranes (PAN10 and PAN400, Sepro) with similar rejection<br />
ratios for separation <strong>of</strong> oil/water emulsions. As ionic liquids can be recycled and<br />
reused without obvious decomposition, the chosen method also demonstrates a<br />
benign pathway to fabricate the cellulose barrier layer for other types <strong>of</strong><br />
membranes.<br />
CARB 135
Discovery <strong>of</strong> 6-deoxy-D-altrose in nature<br />
Masakuni Tako (1) , tako@eve.u-ryukyu.ac.jp, 1 Senbaru, Nishihara Okinawa 903-<br />
0213, Japan ; Takuya Yogi (1) ; Ken Izumori (2) ; Hideharu Ishida (3) ; Makoto Kiso (3) .<br />
(1) Department <strong>of</strong> Sub-tropical Bioscience and Biotechnology, University <strong>of</strong> the<br />
Ryukyus, Nishihara Okinawa 903-0213, Japan (2) Rare Sugar Research Center,<br />
Kagawa University, Miki Kagawa 761-0761, Japan (3) Department <strong>of</strong> Applied<br />
Biochemistry, Gifu University, Gifu Gifu 501-1112, Japan<br />
A novel acetylfucoidan has been isolated [1,2] and covered by patent [3] from<br />
commercially cultured Cladosiphon okamuranus. The acetylfucoidan has now<br />
been produced on the industrial scale and used commercially as the supplement<br />
<strong>of</strong> health foods and cosmetics additives in Japan [4]. We report here the isolation<br />
<strong>of</strong> a novel polysaccharide which substituted 6-deoxy-D-altrose from fruiting<br />
bodies <strong>of</strong> a edible mushroom (Lactarius akahatsu). The paper is the second<br />
report to identify 6-deoxy-D-altrose in nature[5].<br />
Fruiting bodies <strong>of</strong> L. akahatsu were collected on January and February, 2008 in<br />
Okinawa, Japan. Dried powdered samples were suspended in distilled water and<br />
stirred at 100 for 3h to extract a polysaccharide. Total carbohydrate and uronic<br />
acid <strong>of</strong> the polysaccharide was 76.5% and 9.8%, respectively. After acid<br />
hydrolysis <strong>of</strong> the polysaccharide, D-Glc, D-Gal, D-GlcUA and unkown sugar was<br />
identified by HPLC and The 6-deoxy sugar residue was detected by 1 H- and 13 C-<br />
NMR spectroscopy <strong>of</strong> the polysaccharide. The 6-deoxy sugar was identified as a<br />
6-deoxy-D- or L-altrose by 1 H- and 13 C-NMR, COSY and HMQC spectra. The<br />
specific rotation <strong>of</strong> the sugar was estimated to be +18.2 degree. Thus, the<br />
unkown sugar was identified as the 6-deoxy-D-altrose. References:[1] M. Tako et<br />
al.: J. Appl. Glycosci., 43, 143-148 (1996).[2] M. Tako et al.: Botanica Marina, 43,<br />
393-398 (2000).[3] M. Tako: Japanese Patent No. 3371124 (2002).[4] M. Tako:<br />
Botanica Marina, 46, 461-465 (2003).[5]M. Tako et al.,15th European<br />
<strong>Carbohydrate</strong> Symposium, July 19-24, 2009, Wien, Austria.
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