Hybridoma · 2000 · 23 citations · 20 references
Life SciencesInfectious DiseasesMolecular BiologyImmunizationMolecular VirologyMonoclonal AntibodiesAntibody CharacterizationAntibody ScreeningHybridoma TechnologyMonoclonal Antibody TechnologyRecombinant AntibodiesViral HepatitisAmino AcidsHybridoma CharacterizationHybridoma-derived AntibodiesHybridoma-derived Recombinant AntibodiesImmunoglobulin G ResponsesIndirect Enzyme-linked Immunosorbent AssayRecombinant Fusion Proteins
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 enzyme-linked 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, K) recognize aa 21-35, while KR1 (IgG2a, K) recognizes aa 35-47 of the preS1. These MAbs immunoprecipitated HBV particles, demonstrating that they bind to native HBV particles.
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Large surface proteins of hepatitis B virus containing the pre-s sequence
K.‐H. Heermann, U Goldmann, W Schwartz et al. · Journal of Virology · 1984 · 820 citations · Full text