Renaturation from urea

ChenHA hzhen at freeuk.com
Thu Mar 30 07:48:34 EST 2000


"Richard P. Grant" wrote:
> 
> I guess this has been asked before, but as an old hand at protein
> purification but new to inclusion bodies, what do people find the best
> way to renature from 6M urea?
>

My immediate reaction is to suggest that you try to find a
way of getting your bugs to express soluble protein. 
Protein refolding can be a painful business, some refold
easily, some don't, and I don't think there is a universal
methods that can guarantee that your protein refolds.  The
best thing perhaps is to look at some reviews:

    Eliana De Bernardez Clark
    Current Opinion in Biotechnology 1998, 9:157-163.

    Hauke Lilie, Elisabeth Schwarz, Rainer Rudolph
    Current Opinion in Biotechnology 1998, 9:497-501.  
 
It would be also useful to have a look at the following:
http://www.st-and.ac.uk/~lv2/

I'm not suggesting that you buy the products, as I've tried
it and it was only partially successful (similar in
effectiveness to refolding in 1M arginine).  There are,
however, some useful ideas about the process of refolding of
protein.  Good luck (and you'll need it!).
 
> Cheers,
> 
> R
> 
> --
> Richard P. Grant MA DPhil
> Structural Studies Group, MRC-LMB
> http://www2.mrc-lmb.cam.ac.uk/personal/rpg/index.html
> Please reply to rpg 'at' mrc-lmb.cam.ac.uk




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