title: Refolding human serum albumin at relatively high protein concentration.
abstract: 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 M 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 microM). Using this protocol it was possible to refold HSA at concentrations in excess of 5 mg.ml-1 whilst maintaining a high recovery of native monomer. These results represent a considerable improvement on established methods of HSA refolding.
authors.affiliation: Delta Biotechnology Ltd, Nottingham, England.
authors.forename: S J
authors.initials: SJ
authors.lastname: Burton
authors.forename: A V
authors.initials: AV
authors.lastname: Quirk
authors.forename: P C
authors.initials: PC
authors.lastname: Wood
mesh_terms: D001663:Bilirubin / Q000097:blood; D003720:Densitometry; D003956:Dialysis; D004591:Electrophoresis, Polyacrylamide Gel; D006801:Humans; D010084:Oxidation-Reduction; D010449:Peptide Mapping; D011485:Protein Binding; D011487:Protein Conformation; D011489:Protein Denaturation; D012709:Serum Albumin; D013438:Sulfhydryl Compounds / Q000097:blood; D014462:Ultrafiltration
chemical_list: D012709:Serum Albumin; D013438:Sulfhydryl Compounds; D001663:Bilirubin