Conformation of group “a” epitope in hepatitis B surface antigen

Abstract

To elucidate structure of group “a” epitope, mouse antibodies that express idiotype monoclonal antibody and anti-idiotype monoclonal antibody against the group-specific “a” determinant were purified by hydroxyapatite column. To obtain hepatitis B surface antigens (HBsAg), HBsAg positive blood was sequencially purified by ammonium sulfate precipitation, hydroxyapatite, sepharose 4B column chromatography and ultracentrifugation. The major protein (p25) and glycoprotein (gp30) of HBsAg were, isolated by concanavalin-A sepharose 4B. The ability of p25-gp30 among the HBsAg to inhibit the idiotype-anti-idiotype reaction was dependent on conformation, since reduced and alkylated p25-gp30 virtually lost their inhibitory capacity when compared to native HBsAg. The data suggest that hepatitis B antigen is a conformational antigen critically dependent upon the disulfide bonds of p25-pg30.