Mouse monoclonal antibodies to hepatitis B virus PreS1 produced after immunization with recombinant PreS1 peptide

We have efficiently generated mouse monoclonal antibodies (MAbs), which bin d specifically to amino acids 21-47 of the preS1 domain of hepatitis B viru s (HBV) by immunizing mice with the preS1 peptide (amino acids, aa 1-56) co njugated to keyhole limpet hemocyanin, Hybridomas were screened by an indir ect enzyme-linked immunosorbent assay (ELISA) using the purified preS1 pept ide as a coated antigen. Eighteen positive hybridomas were selected and sub jected to isotyping, Of these, 5 clones secreted immunoglobulin G (IgG) and 13 clones secreted IgM, Four (KR1, KR2, KR3, and KR4) of the 5 IgG MAbs bo und to preS1 peptide (aa 21-47), Epitope mapping using bacterially expresse d GST fusion proteins revealed that three clones (KR2, KR3, KR4) (IgG1, kap pa) recognize aa 21-35, while KR1 (IgG2a, kappa) recognizes aa 35-47 of the preS1, These MAbs immunoprecipitated HBV particles, demonstrating that the y bind to native HBV particles.