Expression of pre-S2 region of hepatitis B surface antigen in Escherichia coli.
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 125I-polymerized human albumin cross-linked with glutaraldehyde but not to 125I-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 of HBsAg to solid-phase polymerized the binding of HBsAg to solid-phase polymerized human albumin.
Yu
M W
MW
Division of Blood and Blood Products, Center for Biologics Evaluation and Research, Food and Drug Administration, Bethesda, MD 20892.
Shih
J W
JW
Bursztyn-Pettegrew
H
H
Byars
N E
NE
Allison
A C
AC
Chan
H W
HW
D000818:Animals; D001483:Base Sequence; D003001:Cloning, Molecular; D004279:DNA, Viral / Q000235:genetics*; D004797:Enzyme-Linked Immunosorbent Assay; D004926:Escherichia coli / Q000235:genetics*; D005260:Female; D006168:Guinea Pigs; D006508:Hepatitis Antibodies / Q000276:immunology; D006514:Hepatitis B Surface Antigens / Q000235:genetics* / Q000276:immunology; D006515:Hepatitis B virus / Q000235:genetics*; D006801:Humans; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D004279:DNA, Viral; D006508:Hepatitis Antibodies; D006514:Hepatitis B Surface Antigens; D011993:Recombinant Fusion Proteins