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  • Size Exclusion C hrom

    atography – Principles and M ethods

    gelifesciences.com GE, the GE Monogram, ÄKTA, Biacore, BioProcess, HiLoad, HiPrep, HiScale, HiTrap, illustra, MicroSpin, MidiTrap, MiniTrap, MultiTrap, Sephacryl, Sephadex, Sepharose, SPARTAN, SpinTrap, Superdex, Superose, Tricorn, UNICORN, and Whatman are trademarks of GE Healthcare Company. Tween is a trademark of Uniqema Americas LLC. All other third-party trademarks are the property of their respective owners. IMAC Sepharose products, Ni Sepharose products and Fe Sepharose products: These products are sold under a license from Sigma-Aldrich under patent number EP 1276716 (Metal chelating compositions) and equivalent patents and patent applications in other countries. © 2000–2018 General Electric Company. All goods and services are sold subject to the terms and conditions of sale of the company within GE Healthcare which supplies them. A copy of these terms and conditions is available on request. Contact your local GE Healthcare representative for the most current information.

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    Size Exclusion Chromatography Principles and Methods

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  • Size exclusion chromatography Principles and Methods

  • 2 18102218 AM

    Contents Symbols, acronyms and terminology used in this handbook ......................................................... 5

    Symbols ............................................................................................................................................................................. 5

    Common acronyms and abbreviations .............................................................................................................. 5

    Chromatography terminology ................................................................................................................................ 7

    Chapter 1 Introduction ............................................................................................................................................11

    Versatility of size exclusion chromatography .............................................................................................. 11

    Purification by size exclusion chromatography .......................................................................................... 12

    Size exclusion chromatography in theory...................................................................................................... 14

    Defining the process ....................................................................................................................................... 14

    Selectivity curves and resin selection ..................................................................................................... 17

    Resolution ............................................................................................................................................................ 19

    Chapter 2 Size exclusion chromatography in practice ......................................................................................21

    General advice ............................................................................................................................................................ 21

    Columns and resin preparation ................................................................................................................. 21

    Scaling up ............................................................................................................................................................ 21

    Sample and buffer preparation ................................................................................................................. 22

    Filtration ................................................................................................................................................................ 22

    Equipment selection ....................................................................................................................................... 23

    Setting column pressure limits ................................................................................................................. 23

    Maintenance of SEC columns ..................................................................................................................... 24

    Optimizing your separation ................................................................................................................................. 24

    Resin selection ................................................................................................................................................... 25

    Sample volume and column dimensions .............................................................................................. 26

    Elution and flow rates .................................................................................................................................... 27

    Sample concentration and viscosity ..............................................................................