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Ion Exchange Chromatography Principles and Methods GE Healthcare GE Imagination at Work at Imagination Ion Exchange Chromatography – Principles and Methods Q, Q GE Healthcare Ion Exchange Chromatography – Principles and Methods www.gelifesciences.com GE, GE monogram, Amersham, ÄKTA, AxiChrom, Biacore, BioProcess, Capto, Cy, CyDye, ECL, ECL Plex, ECL Select, FineLINE, HiPrep, HiScale, HiScreen, HiTrap, Hybond, MabSelect SuRe, MiniBeads, Mini Q, Mini S, MiniTrap, MonoBeads, Mono Q, Mono S, PhastGel, PhastSystem, PreDictor, PrimeView, Protran, ReadyToProcess, RESOURCE, Sephadex, Sepharose, SOURCE, SPARTAN, Superdex, Tricorn, UNICORN, and Whatman are trademarks of General Electric Company. Coomassie is a trademark of Thermo Fisher Scientific LLC, Gyrolab is a trademark of Gyros AB. RoboColumn is a trademark of Atoll GmbH. Triton is a trademark of Union Carbide Chemicals and Plastic Company Inc. Tween is a trademark of Croda Group of Companies. 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Methods 11000421 AC 1 Contents Introduction.........................................................................................................................................7 Symbols ............................................................................................................................................................................... 8 Common acronyms and abbreviations ............................................................................................................... 8 Chapter 1 Principles of ion exchange ............................................................................................................. 11 Net surface charge and pH .....................................................................................................................................11 Steps in an IEX separation .......................................................................................................................................12 Equilibration .............................................................................................................................................................12 Sample application and wash ........................................................................................................................12 Elution .........................................................................................................................................................................12 Regeneration .........................................................................................................................................................12 Resolution .........................................................................................................................................................................14 Efficiency ...................................................................................................................................................................15 Selectivity ..................................................................................................................................................................17 Components of IEX media ........................................................................................................................................21 Matrix ..........................................................................................................................................................................21 Functional groups .................................................................................................................................................23 Binding capacity and recovery ..............................................................................................................................24 Chromatofocusing........................................................................................................................................................25 Chapter 2 Ion exchange in practice ............................................................................................................... 27 Introduction .....................................................................................................................................................................27 Selecting chromatography media .......................................................................................................................27
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