Premium
Expression of pre‐S2 region of hepatitis B surface antigen in Escherichia coli
Author(s) -
Yu M. W.,
Shih J. W.K.,
BursztynPettegrew H.,
Byars N. E.,
Allison A. C.,
Chan H. W.
Publication year - 1990
Publication title -
journal of medical virology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.782
H-Index - 121
eISSN - 1096-9071
pISSN - 0146-6615
DOI - 10.1002/jmv.1890300103
Subject(s) - hbsag , recombinant dna , antiserum , microbiology and biotechnology , virology , fusion protein , antigen , yersinia pestis , biology , western blot , hepatitis b virus , human serum albumin , virus , biochemistry , gene , immunology , virulence
We constructed a recombinant plasmid that can express the entire pre‐S2 sequence of hepatitis B surface antigen (HBsAg) as a fusion protein in E. coli . The hybrid protein, which comprises the bacterial TrpLE protein and the pre‐S2 sequence, was the prominent protein that was found in cell extracts. As determined by immune blot analysis, this protein reacted with human HBV convalescent sera, as well as with sera from animals immunized with either purified HBsAg or isolated polypeptides containing pre‐S2. It bound specifically to 125 I‐polymerized human albumin cross‐linked with glutaraldehyde but not to 125 I‐monomeric human albumin. A novel adjuvant formulation was used in place of Freund's adjuvant to immunize guinea pigs with the recombinant product. The antisera obtained from serial bleedings were found to react with HBsAg of both d and y subtypes. These antisera were also shown to react solely with HBsAg polypeptides which contain the pre‐S2 sequence and to block the binding of HBsAg to solid‐phase polymerized human albumin.