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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