Refolding issue a cysteine rich peptide

[Frank Jeanplong]

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]

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]

i can suggest a good paper about refolding of recombinant proteins

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