Ion Exchange Chromatography - ... Protein Purification Handbook 18-1132-29 Ion Exchange...

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  • 18-1114-21

    Edition AB

    Ion Exchange Chromatography Principles and Methods

    Ion Exchange Chrom atography – Principles and M

    ethods

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    www.amershambiosciences.com (Click on Chromatography)

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    Sephadex, Sephacel, Sepharose, STREAMLINE, HiLoad, HiTrap, MonoBeads, SOURCE, RESOURCE, FPLC, FPLCdirector, UNICORN, SMART, OligoPilot II, FineLine, BPG, BioPilot, BioProcess, PhastSystem, PhastGel and Drop Design are trademarks of Amersham Biosciences Limited.

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    Protein Purification Handbook 18-1132-29

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  • Ion Exchange Chromatography

    Principles and Methods

    ISBN 91 970490-3-4

  • 1. Introduction ............................................................................................9

    2. Ion exchange chromatography .........................................................10 The theory of ion exchange.................................................................10

    The matrix.....................................................................................11 Charged groups .............................................................................13 Resolution in ion exchange chromatography .................................13

    Capacity factor ........................................................................15 Efficiency.................................................................................16 Selectivity.................................................................................17

    Capacity ........................................................................................18

    3. Product Guide .......................................................................................20 MonoBeads ...................................................................................20 MiniBeads .....................................................................................20 SOURCE .......................................................................................20 Sepharose High Performance ion exchangers .................................21 Sepharose Fast Flow ion exchangers ..............................................21 Sepharose Big Beads ion exchangers...............................................21 STREAMLINE ion exchangers ......................................................21 DEAE Sepharose CL-6B and CM Sepharose CL-6B .......................22 DEAE Sephacel..............................................................................22 Sephadex ion exchangers ...............................................................22 Bulk quantities...............................................................................22 Equipment.....................................................................................22

    4. MonoBeads and MiniBeads ..............................................................23 MonoBeads.....................................................................................23

    Properties ......................................................................................24 Chemical stability ....................................................................24 Physical stability ......................................................................25 Flow rate..................................................................................27 Capacity ..................................................................................27 Recovery..................................................................................28 Reproducibility ........................................................................29

    Availability ....................................................................................30 MiniBeads .......................................................................................30

    Properties ......................................................................................31 Chemical stability ....................................................................31 Physical stability ......................................................................32 Reproducibility ........................................................................33

    Availability ....................................................................................33

    Contents

    2

  • 5. SOURCE ................................................................................................34 Properties ......................................................................................37

    Chemical stability ....................................................................37 Flow rate..................................................................................38 Capacity ..................................................................................40 Recovery..................................................................................40 Reproducibility ........................................................................41

    Availability ....................................................................................41

    6. Sepharose based ion exchangers .......................................................42 Chemical stability ..........................................................................42 Physical stability ............................................................................43

    Sepharose High Performance ion exchangers.........................43 Properties ......................................................................................44

    Physical stability ......................................................................44 Capacity ..................................................................................44 Flow rate..................................................................................46

    Availability ....................................................................................46 Sepharose Fast Flow ion exchangers ........................................46

    Properties ......................................................................................47 Physical stability ......................................................................47 Capacity ..................................................................................47 Flow rate..................................................................................49

    Availability ....................................................................................50 Sepharose Big Beads ion exchangers.........................................50

    Properties ......................................................................................51 Physical properties ...................................................................51 Capacity ..................................................................................51 Flow rate..................................................................................51

    Availability ....................................................................................51 STREAMLINE SP and STREAMLINE DEAE ......................52

    Properties ......................................................................................53 Physical stability ......................................................................53 Capacity ..................................................................................53

    Availability ....................................................................................54 DEAE Sepharose CL-6B and CM Sepharose CL-6B.............54

    Properties .....................