BioHPLC Column Selection Guide Cover - Agilent Technologies

More documents

Recommendations

Info

ion-exchange Chromatography Column Specifications Dimensions 5.2 mm x 4.95 mm Column volume 100 µL Maximum pressure 150 bar (15 MPa, 2200 psi) Temperature min/max Working: 4-40 °C Storage: 4-30 °C Recommended pH Working range: 2-13 Cleaning-in-place: 1-14 Materials of construction Hardware: Stainless steel Packing: poly (glycidyl methacrylate-co-ethylene dimethacrylate) highly porous monolith Color ring identifier Bio-Monolith QA: Blue Bio-Monolith DEAE: Green Bio-Monolith SO3: Red Shelf life/expiration date SO3, QA, DEAE: 24-36 months 2 www.agilent.com/chem/biohplc Tips & Tools agilent Bio-Monolith Protein a affinity columns are found on page 9.
Baseline expansion of a separation of protein standards Column: Agilent Bio-Monolith CM15, 5.5 x 15 mm Mobile Phase: A: 10 mM Na2HPO4, pH 6.0 B: A + 0.5 M NaCl or just 0.5 M Na2HPO4, pH 6.0 Flow Rate: 2 mL/min Gradient: 0.5 min hold with mobile phase A followed by a linear gradient to 45% B in 15 min (elapsed time 15.5 min); then 60% B at 15.6 min continued to 20 min. Column flushed with 100% B for 15 min before re-equilibration for the next run. pH Gradient: A: 5 mM Na2HPO4, buffer pH 5.5 and B: 40 mM NA2HPO4 (not buffered, pH 8.9). 2% B/min at 1 mL/min for 15 min, followed by a column wash with 90% B for 5 min. Detector: UV at 220 nm Sample: One mg each/mL in mobile phase A. 1. RNAse from bovine pancreas (pI 9.6) 2. Cytochrome c from bovine heart (pI 10.37-10.8) 3. Lysozyme from chicken egg (pI 11.35) (0.5 mg) mAU 14 12 10 8 6 4 mAU 14 12 10 8 6 4 ion-exchange Chromatography 4 6 8 10 12 14 16 18 min 4 4.483 4.968 4.284 4.520 4.688 4.945 5.653 6.357 6.834 1 2 3 A with NaCl gradient 7.832 6.482 7.059 7.604 8.162 9.072 9.271 10.335 12.170 B with Na 2HPO 4 gradient 6 8 10 12 14 16 18 min Instrument: Agilent 1200 SL with diode array detector B shows a better resolution of protein contaminants. 16.410 16.901 UHPLC/HPLC 3
Page 1 and 2:
Agilent BioHPLC Column Selection Gu
Page 3 and 4:
BIOCOLUMN SELECTION GUIDELINES From
Page 5 and 6:
Column Selection Flow Chart Biocolu
Page 7 and 8:
Protein Column Selection Guide Appl
Page 9 and 10:
Separation of charge variants of hu
Page 11 and 12:
Increased resolution for peptide ma
Page 13 and 14: Preparative scale purification of L
Page 15 and 16: DNA and RNA Oligonucleotide Column
Page 17 and 18: Broad Distribution Biomolecules Car
Page 19 and 20: ReveRseD-PHase HPLC Confidently per
Page 21 and 22: Sterically Protected 300StableBond
Page 23 and 24: Improved reproducibility of monoclo
Page 25 and 26: Short-chain ZORBAX 300SB-C3 is stab
Page 27 and 28: ZORBAX 300Å StableBond Hardware De
Page 29 and 30: LC/MS analysis of angiotensin on Ex
Page 31 and 32: Tips & Tools ZORBAX 300Å Extend-C1
Page 33 and 34: Poroshell 300 columns separate prot
Page 35 and 36: Monoclonal IgG1 chains: Separation
Page 37 and 38: Poroshell 120 • 120Å pore size f
Page 39 and 40: 0 µm 0 µm 0 µm A B C Scanning el
Page 41 and 42: Exploiting chemical stability - TFA
Page 43 and 44: Temperature as a tool to enhance ma
Page 45 and 46: ZORBaX amino acid analysis (aaa) Co
Page 47 and 48: High resolution of 24 amino acids u
Page 49 and 50: Ion-Exchange Column Selection Appli
Page 51 and 52: Consistent ion-exchange MAb separat
Page 53 and 54: Column Specifications Tips & Tools
Page 55 and 56: Weak cation-exchange chromatography
Page 57 and 58: PL-SAX Strong Anion-Exchange Column
Page 59 and 60: Analysis of representative whey pro
Page 61 and 62: Standard protein separation Column:
Page 63: Agilent Bio-Monolith HPLC Column Se
Page 67 and 68: size exclusion Chromatography (seC)
Page 69 and 70: Column Specifications Agilent Bio S
Page 71 and 72: Comparison of Agilent Bio SEC-3 and
Page 73 and 74: Column Length size exclusion Chroma
Page 75 and 76: Column Specifications Agilent Bio S
Page 77 and 78: Calibration curves - Bio SEC-5 Colu
Page 79 and 80: ProSEC 300S • Mechanically robust
Page 81 and 82: Overlay of UV and light scattering
Page 83 and 84: Retention volume versus log (MW) fo
Page 85 and 86: Agilent PL aquagel-OH SEC Columns f
Page 87 and 88: Heparin Column: 2 x PL aquagel-OH 3
Page 89 and 90: Polyethylene Glycol/Oxide standards
Page 91 and 92: aFFiniTY CHROMaTOGRaPHY Agilent Bio
Page 93 and 94: Rapid human polyclonal IgG quantifi
Page 95 and 96: Multiple Affinity Removal System af
Page 97 and 98: Illustration of high abundance prot
Page 99 and 100: mRP-C18 High-Recovery Protein Colum
Page 101 and 102: MeTHOD DeveLOPMenT ZORBAX Column Me
Page 103 and 104: Reversed-Phase LC/MS Methods Method
Page 105 and 106: Method Development Ionic Strength:
Page 107 and 108: Method Development After the initia
Page 109 and 110: Tips & Tools Agilent offers a varie
Page 111 and 112: Human serum: Low abundance protein
Page 113 and 114: 2-D LC/MS Analyses Using ZORBAX Cap
Page 115 and 116:
ZORBAX Bio-SCX Series II Capillary
Page 117 and 118:
ZORBAX HPLC Capillary Columns (glas
Page 119 and 120:
Microbore HPLC for sensitive peptid
Page 121 and 122:
BioPharmaceutical Lifecycle Reverse
Page 123 and 124:
ZORBAX 300Å StableBond ZORBAX Prep
Page 125 and 126:
PLRP-S Prep to Process Application
Page 127 and 128:
Prep to Process PLRP-S Size (mm) Pa
Page 129 and 130:
PL-SAX AND PL-SCX FOR PREP TO PROCE
Page 131 and 132:
Prep to Process PL-SAX and PL-SCX D
Page 133 and 134:
Crude peptide screen Column: VariTi
Page 135 and 136:
CARTRIDGE COLUMN SYSTEMS Cartridge
Page 137 and 138:
USP DESIGNATIONS - BIOHPLC COLUMNS
Page 139 and 140:
BioHPLC Columns Literature Title Ag
Page 141 and 142:
BioHPLC Columns Literature Title An
Page 143 and 144:
BioHPLC Columns Literature Title Co
Page 145 and 146:
BIOPHARM DEFINITIONS A active start
Page 147 and 148:
dimer A polymer made up of two iden
Page 149 and 150:
K-L ligase An enzyme that causes mo
Page 151 and 152:
S secondary structure In proteins,
Page 153 and 154:
Poroshell 300 Citations Matilainen,
Page 155 and 156:
Agilent 1260 Infinity Bio-inert Qua
show all

BioHPLC Column Selection Guide Cover - Agilent Technologies

You also want an ePaper? Increase the reach of your titles

Delete template?

Save as template?