Short Communication: Mouse Monoclonal Antibodies to Hepatitis B Virus PreS1 Produced After Immunization with Recombinant PreS1 Peptide

Abstract

We have efficiently generated mouse monoclonal antibodies (MAbs), which bind specifically to amino acids 21–47 of the preS1 domain of hepatitis B virus (HBV) by immunizing mice with the preS1 peptide (amino acids, aa 1–56) conjugated to keyhole limpet hemocyanin. Hybridomas were screened by an indirect enzymelinked immunosorbent assay (ELISA) using the purified preS1 peptide as a coated antigen. Eighteen positive hybridomas were selected and subjected 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 bound to preS1 peptide (aa 21–47). Epitope mapping using bacterially expressed GST fusion proteins revealed that three clones (KR2, KR3, KR4) (IgG1, κ) recognize aa 21–35, while KR1 (IgG2a, κ) recognizes aa 35–47 of the preS1. These MAbs immunoprecipitated HBV particles, demonstrating that they bind to native HBV particles.