Monoclonal antibodies against 24 kDa surface antigen of hepatitis B viruses

Hepatitis B virus (HBV) infection is one of the major public health problems worldwide. Effective control of HBV transmission in areas of high and intermediate endemicity would not be possible without vaccination of the vulnerable group. The diagnosis of acute and chronic hepatitis B infection is based on the detection of hepatitis B surface antigen (HBsAg). In this study, we prepared nine hybridomas that produced monoclonal antibodies specific for HBsAg. BALB/c mice were immunized with 24 kDa HBsAg and the immune spleen cells were fused with SP2/0 myeloma cell line. We obtained seven IgG1, one IgM, and one IgG2b positive clones. The stable hybrids were sub-cloned and ascitic fluid was prepared in the BALB/c mice. After antibody purification by protein G, the affinity column was prepared and the pure 24 kDa HBsAg from cell extract was eluted from the column. Western blot analysis showed that all monoclonal antibodies are specific for 24 kDa antigen. Since 24 kDa HBsAg is used for vaccination against hepatitis, these monoclonal antibodies are the best candidate for the isolation and purification of recombinant HBsAg from yeast expression vector by affinity column to be used for vaccination.