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Size exclusion chromatography for 23 and 20.5KDa? - protein purification (Sep/19/2007 )

Hi, I have two proteins that are 23KDa and 20.5KDa in size. Is it possible to seperate them using a Superdex 200 10/300 GL column in FPLC? Anyone has experiences with it, or columns to recommend? Thanks guys!


you probably won't see any significant separation with superdex 200. you may be able to get a good enough separation with superdex 75 (if you keep your injection volume low and not too much protein).


Agree with Mdfenko. Superdex 75 will be better ( because Kav for this Mw range is in optimal range ( 0.4-0.5) or as alternative may be Superose 12, but in general it will be better to change method of separation ( by charge IEC chromatography) or by hydrofobicity ( HIC chromatography). I think it will be
a short way to separation of closed Mw proteins.


Hey guys.. thx for the input - appreciate it.