fulltext.study @t Gmail

In vitro-refolding of a single-chain Fv fragment in the presence of heteroaromatic thiols

Paper ID Volume ID Publish Year Pages File Format Full-Text
24770 43536 2008 4 PDF Available
Title
In vitro-refolding of a single-chain Fv fragment in the presence of heteroaromatic thiols
Abstract

The aim of the present work was to explore the use of heteroaromatic thiol compounds, namely derivatives of pyridine and pyrimidine, as redox reagents for the in vitro-refolding of a recombinantly expressed single-chain Fv fragment (scFvOx). The mixed disulfide of scFvOx with glutathione was used as a starting material, while reduced glutathione, 4-mercaptopyridine, 2-mercaptopyrimidine, 2-mercaptopyridine N-oxide, and the mercaptobenzene derivative thiosalicylic acid, respectively, served as catalysts for the formation of native disulfide bonds during renaturation. In contrast to thiosalicylic acid, and despite their significantly lower thiol pKa values, none of the heteroaromatic thiol compounds accelerated the apparent kinetics of in vitro-refolding compared to the naturally occurring peptide glutathione. However, significantly improved renaturation yields were observed in the presence of 4-mercaptopyridine and 2-mercaptopyrimidine, demonstrating the usefulness of aromatic thiol compounds as reagents for the in vitro-refolding of antibody fragments.

Keywords
In vitro-refolding; Redox buffer; Antibody fragments; Mixed disulfide; Aromatic thiol compounds
First Page Preview
In vitro-refolding of a single-chain Fv fragment in the presence of heteroaromatic thiols
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 134, Issues 3–4, 30 April 2008, Pages 218–221
Authors
, , ,
Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering