GST fusions - removal of proteases

Charlie Bond charlieb at biochem.usyd.edu.au
Mon Aug 4 01:42:27 EST 1997


Sebastian Springer wrote:
> 
> Hi -
> 
> I am trying to find a way to effectively remove thrombin or factor X
> from eluates of cleavage reactions with GST fusions on glutathione
> sepharose. The eluate has two proteins in it, my protein of interest
> and the protease. Do these proteases have any special properties that
> would make their removal easy (e.g. binding properties to some ion
> exchanger)? Who hs developed protocols already?

Sundaram and Brandsma [(J. virological methods) 57 (1996) 61-70] report
the successful use of a benzamidine Sepharose 6B column. We haven't
tried it yet. A previous poster also mentioned anti-thrombin agarose.

Cheers,
Charlie



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