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-   -   Refolding issue a cysteine rich peptide (http://www.molecularstation.com/forum/recombinant-protein-forum/2908-refolding-issue-cysteine-rich-peptide.html)

Frank Jeanplong 01-13-2008 09:55 PM

Refolding issue a cysteine rich peptide
 
I need to refold a cyteine rich 100 odd residue peptide expressed in E.coli. I can get high purity from inclusion bodies and passing it through a Ni-NTA column in 6M guanidium chloride but it aggregates in dialysis. I tried a few different dial cond. but I lost the majority of recombinant protein. I would need a systematic optimization starategy to get the most native/bioactive peptide at the end. Thanks for your time.

ednot 12-22-2008 04:55 PM

Re: Refolding issue a cysteine rich peptide
 
Hello
Try an on-column refolding on imac.

I suggest the following additives
Gluthatione Reduced / Ox
L-Arginine
Glycerol
Mannitol
Triton X100
CuSo4 ( not in IMAC, but in dilution)


check the refold databse


regards.

ed

trypsin 03-04-2009 09:04 PM

Re: Refolding issue a cysteine rich peptide
 
i can suggest a good paper about refolding of recombinant proteins

"In vitro folding of inclusion body proteins" Rainer Rudolph and Hauke Lilie (Review)


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