Purification of GST fusion proteins

K Jolley bspkj at bath.ac.uk
Mon May 1 10:45:24 EST 1995

>>> Our GST fusion protein (approx. 100 kDa) does not bind to Pharmacia Glutathione Sepa
>>> Sepharose under standard conditions. 
>>Is it soluble or is it going into inclusion bodies? 
> I also encountered the same kind of problem of my GST fusion protein, 
>and I was very sure thta it is soluble (confirmed by immunodetection).  
>The fusion protein binds to glutarthion-beads (Sigma) in a very low 
>efficiency manner, and most of the fusion proteins would be washed from 
>the beads with only on wash.
>I-Jen Huang
>Gr8bn at cc.usu.edu
I had a similar problem with an 86kDa fusion. I've since found out that this
is a fuction of the molecular weight of the protein, binding efficiency
diminishes greatly with increasing size. For further details see
Frangioni @ Neel (1993), Anal. Biochem., 210, 179-187.

	Keith Jolley
*Keith Jolley			    * Tel (Work): (01225) 826826 x5134 *
*School of Biology and Biochemistry *	  (Home): (01225) 835259       *	
*University of Bath, UK		    *   Internet: bspkj at bath.ac.uk     *

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