Open AccessRefolding human serum albumin at relatively high protein concentration
Author(s) -
BURTON Steven J.,
QUIRK Alan V.,
WOOD P. Carolyn
Publication year - 1989
Publication title -
european journal of biochemistry
Language(s) - English
Resource type - Journals
eISSN - 1432-1033
pISSN - 0014-2956
DOI - 10.1111/j.1432-1033.1989.tb14564.x
Subject(s) - chemistry , human serum albumin , urea , dialysis , monomer , chromatography , yield (engineering) , albumin , dilution , serum albumin , sodium , bovine serum albumin , biochemistry , organic chemistry , polymer , metallurgy , thermodynamics , medicine , materials science , physics
The conditions for refolding reduced and denatured human serum albumin (HSA) were investigated with a view to maximising the yield of native monomeric albumin. Refolding by dialysis was found to be preferable to dilution as a means of chaotrope (urea) and reductant (2‐mercaptoethanol) removal. Dialysis of denatured HSA solutions containing 4–8 urea and 14 mM 2‐mercaptoethanol at pH 10.0 was found to be optimal for HSA refolding. The yield of monomeric HSA was maximal (94%) for dialysis in the presence of EDTA (1 mM) and sodium palmitate (20 μM). Using this protocol it was possible to refold HSA at concentrations in excess of 5 mg · m1 −1 whilst maintaining a high recovery of native monomer. These results represent a considerable improvement on established methods of HSA refolding.