BioHPLC Column Selection Guide Cover - Agilent Technologies

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Ion-Exchange Chromatography Agilent Bio IEX HPLC Columns, PEEK Size (mm) Particle Size (µm) Pressure Limit 54 www.agilent.com/chem/biohplc Bio SCX Part No. Bio WCX Part No. Bio SAX Part No. Bio WAX Part No. 4.6 x 250 10 275 bar/4000 5190-2435 5190-2455 5190-2475 5190-2495 4.6 x 50, Guard 10 275 bar, 4000 psi 5190-2436 5190-2456 5190-2476 5190-2496 4.6 x 250 5 400 bar, 5800 psi 5190-2427 5190-2447 5190-2467 5190-2487 4.6 x 50, Guard 5 400 bar, 5800 psi 5190-2428 5190-2448 5190-2468 5190-2488 2.1 x 250 10 275 bar, 4000 psi 5190-2439 5190-2459 5190-2479 5190-2499 2.1 x 50, Guard 10 275 bar, 4000 psi 5190-2440 5190-2460 5190-2480 5190-2500 2.1 x 250 5 400 bar, 5800 psi 5190-2431 5190-2451 5190-2471 5190-2491 2.1 x 50, Guard 5 400 bar, 5800 psi 5190-2432 5190-2452 5190-2472 5190-2492 Agilent Bio IEX HPLC Columns, Stainless Steel Size (mm) Particle Size (µm) Pressure Limit Bio SCX Part No. Bio WCX Part No. Bio SAX Part No. Bio WAX Part No. 4.6 x 250 10 275 bar, 4000 psi 5190-2433 5190-2453 5190-2473 5190-2493 4.6 x 250 5 413 bar, 6000 psi 5190-2425 5190-2445 5190-2465 5190-2485 4.6 x 50 3 551 bar, 8000 psi 5190-2423 5190-2443 5190-2463 5190-2483 4.6 x 50 1.7 600 bar, 10000 psi 5190-2421 5190-2441 5190-2461 5190-2481 4.0 x 10, Guard 10 275 bar, 4000 psi 5190-2434 5190-2454 5190-2474 5190-2494 4.0 x 10, Guard 5 413 bar, 6000 psi 5190-2426 5190-2446 5190-2466 5190-2486 4.0 x 10, Guard 3 600 bar, 10000 psi 5190-2424 5190-2444 5190-2464 5190-2484 4.0 x 10, Guard 1.7 275 bar, 4000 psi 5190-2422 5190-2442 5190-2462 5190-2482 Tips & Tools We offer a variety of e-Seminars and on-site training to help you be a more effective chromatographer, at www.agilent.com/chem/education
PL-SAX Strong Anion-Exchange Columns ion-exchange Chromatography • Small particles deliver excellent chromatographic performance • Wide range of particle sizes and 2 pore sizes for flexible analysis to scale-up purification • Exceptional stability for long column lifetime PL-SAX -N(CH 3) 3 + is ideal for the anion-exchange HPLC separations of proteins, peptides and deprotected synthetic oligonucleotides under denaturing conditions. The strong anion-exchange functionality, covalently linked to a chemically stable fully porous polymer, extends the operating pH range. In addition, the anion-exchange capacity is independent of pH. For synthetic oligonucleotides, separations using denaturing conditions of temperature, organic solvent, and high pH are all possible. PL-SAX delivers improved chromatography for self-complementary or G-rich sequences that may associate to form aggregates or hairpin structures. The 5 µm material provides high efficiency separations of n and n-1 sequences. A wide range of particle sizes and column geometries permits analysis scale-up to purification. The strong anion-exchange functionality provides a material with exceptional chemical and thermal stability, even with sodium hydroxide eluents, leading to long column lifetime. Column Specifications Bonded Phase ID Particle Size Pore Size pH Stability Operating Temperature Limit Strong Anion-Exchange 2.1, 4.6, 5, 8, 10 1000Å 1-14 80 ºC 7.5, 25, 50 and 100 and 30 and 4000Å UHPLC/HPLC 55
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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: Weak cation-exchange chromatography
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
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
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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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