Immunochemistry of hepatitis B surface antigen (HBsAg): preparation and characterization of antibodies to the constituent polypeptides

Abstract

The structural polypeptides of HBsAg were shown to be immunogenic in guinea pigs. Purified 22-nm forms of the ad and any subtypes of HBsAg were solubilized under reducing conditions and electrophoresed in SDS-polyacrylamide gels. Individual polypeptides isolated from both HBsAg/ad and HBsAg/ay subtypes were used to hyperimmunize guinea pigs using Freund's complete adjuvant. All animals produced specific antibodies against native HBsAg as determined by complement fixation, passive hemagglutination, and double-antibody radioimmunoprecipitation assays. Each polypeptide contained within its structure the group-specific HBsAg determinant, a. Equilibrium competitive inhibition studies were conducted to determine the relative affinities of antisera produced against the major HBsAg polypeptides P-1, P-2, and P-6 (23,000, 29,000, and 72,000 daltons, respectively).