why do one should use urea or guanidium HcL in invitro refolding of recombinant therapeutic protein ? ?
In our lab we are refolding recombinant Human growth hormone from E.Coli . in that urea and Gunadium Hcl is being used why do one should use urea or guanidium HcL in invitro refolding of recombinant therapeutic protein ? can any one give the exact molarity of Urea or Gunadium Hcl to be used for refolding ? which of the following either urea or Gunadium Hcl is best for refolding ?
One Response
Guru
05 Mar 2010
The basis of renaturation acheived by trition is that it provides a hydrophobic environment to shadow the pockets for the hydrophobic core of the protein. In the absence of a hydrophobic environment the protein may undergo collapse and form a pellet or aggregate.
efolding occurs when you changed from denaturing environment into the aqueous environment .
usually when u refold proteins. anywhere from 10 – 20% of the dentaured proteins is refolded with actvity
refolding or renaturing occured when you washes the gels in 0.5% Triton X-100 in 40 mM Tris-HCl pH 6.0.