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.............................................................................................................................................................................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 ....................................................................................................... 29 Buffer composition .......................................................................................................................................... 31 System configuration ..................................................................................................................................... 33 Troubleshooting .......................................................................................................................................................... 35 Reference ............................................................................................................................................................. 38 Chapter 3 Considerations for analytical SEC ......................................................................................................39