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? 
>
>>cheers,
>>John
> 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.

Regards,
	Keith Jolley
---
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