BioHPLC Column Selection Guide Cover - Agilent Technologies

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Reversed-Phase HPLC Long life at high pH with 300Extend-C18 Column: ZORBAX Extend-C18 773450-902 4.6 x 150 mm, 5 µm Mobile Phase: 20% 20 mM NH4OH, pH 10.5 80% Methanol Flow Rate: 1.5 mL/min Temperature: Aging 24 °C Tests 40 °C Each 10,000 column volume is approximately one working month. Use ZORBAX Extend-C18 for alternate selectivity at high pH Column: ZORBAX Extend-C18 773700-902 2.1 x 150 mm, 5 µm Mobile Phase: Flow Rate: 0.25 mL/min Gradient: 5-60% B in 20 min Temperature: 25 °C A: 0.1% TFA in Water B: 0.085% TFA in 80% ACN A: 20 mM NH4OH in Water B: 20 mM NH4OH in 80% ACN MS Conditions: Pos. Ion ESI-Vf 70V, Vcap 4.5 kV N2 – 35 psi, 12 L/min, 300 °C 4 µL (50 ng each peptide) The Extend column can be used for high pH separations of peptides. At high and low pH, very different selectivity can result. Just by changing pH, a complimentary method can be developed and it is possible to determine if all peaks are resolved. The Extend column can be used at high and low pH, so the complimentary separation can be investigated with one column. Better MS sensitivity for this sample is also achieved at high pH. 2 www.agilent.com/chem/biohplc k Values 8 6 4 2 0 0 10,000 20,000 30,000 40,000 50,000 Column Volumes of Mobile Phase 3E7 2.5E7 2E7 1.5E7 1E7 .5E7 3E7 2.5E7 2E7 1.5E7 1E7 A B 4-Chloro-1-nitrobenzene Trimipramine LHL LHG LHG LC30001 .5E7 0 0 2 4 6 8 10 12 14 16 18 20 Time (min) LLL LHL LLVF LLL-NH2 LLL 0 0 2 4 6 8 10 12 14 16 18 20 LLVF LLL-NH 2 LCBP017
Tips & Tools ZORBAX 300Å Extend-C18 Reversed-Phase HPLC Hardware Description Size (mm) Particle Size (µm) Part No. Analytical 4.6 x 250 5 770995-902 Analytical 4.6 x 150 5 773995-902 Rapid Resolution 4.6 x 150 3.5 763973-902 Rapid Resolution 4.6 x 100 3.5 761973-902 Rapid Resolution 4.6 x 50 3.5 765973-902 Narrow Bore RR 2.1 x 150 3.5 763750-902 Narrow Bore RR 2.1 x 100 3.5 761775-902 Narrow Bore RR 2.1 x 50 3.5 765750-902 Guard Cartridge, 4/pk 4.6 x 12.5 5 820950-932 Guard Cartridge, 4/pk 2.1 x 12.5 5 821125-932 Guard Hardware Kit 820999-901 Capillary Glass-lined Columns Capillary RR 0.3 x 150 3.5 5065-4464 Capillary RR 0.3 x 100 3.5 5065-4465 Capillary RR 0.3 x 75 3.5 5065-4466 Capillary RR 0.3 x 50 3.5 5065-4467 Guard columns and filters help protect your column and instruments from particulates that can cause blockages, which increase system pressure and negatively impact performance. Learn how to fix this common problem at www.agilent.com/chem/troubleshooting. UHPLC/HPLC 29
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: LC/MS analysis of angiotensin on Ex
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 and 64: Agilent Bio-Monolith HPLC Column Se
Page 65 and 66: Baseline expansion of a separation
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
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Multiple Affinity Removal System af
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Illustration of high abundance prot
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mRP-C18 High-Recovery Protein Colum
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MeTHOD DeveLOPMenT ZORBAX Column Me
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Reversed-Phase LC/MS Methods Method
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Method Development Ionic Strength:
Page 107 and 108:
Method Development After the initia
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Tips & Tools Agilent offers a varie
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Human serum: Low abundance protein
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2-D LC/MS Analyses Using ZORBAX Cap
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ZORBAX Bio-SCX Series II Capillary
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ZORBAX HPLC Capillary Columns (glas
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Microbore HPLC for sensitive peptid
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BioPharmaceutical Lifecycle Reverse
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ZORBAX 300Å StableBond ZORBAX Prep
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PLRP-S Prep to Process Application
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Prep to Process PLRP-S Size (mm) Pa
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PL-SAX AND PL-SCX FOR PREP TO PROCE
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Prep to Process PL-SAX and PL-SCX D
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Crude peptide screen Column: VariTi
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CARTRIDGE COLUMN SYSTEMS Cartridge
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USP DESIGNATIONS - BIOHPLC COLUMNS
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BioHPLC Columns Literature Title Ag
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BioHPLC Columns Literature Title An
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BioHPLC Columns Literature Title Co
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BIOPHARM DEFINITIONS A active start
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dimer A polymer made up of two iden
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K-L ligase An enzyme that causes mo
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S secondary structure In proteins,
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Poroshell 300 Citations Matilainen,
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Agilent 1260 Infinity Bio-inert Qua
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BioHPLC Column Selection Guide Cover - Agilent Technologies

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