Mapping of Monoclonal Antibody Epitopes of the Rabies Virus P Protein

J Gen Virol. 1997 Jan;78 ( Pt 1):119-24. doi: 10.1099/0022-1317-78-1-119.

Abstract

Thirty-six monoclonal antibodies (MAbs) specific for the rabies virus P phosphoprotein were obtained from mice immunized with recombinant P (PV strain) produced in E. coli. All MAbs reacted against the corresponding rabies virus protein by ELISA and by Western blot analysis and revealed the presence of cytoplasmic inclusions in rabies virus infected cells. The epitopes of seven MAbs were mapped by testing their reactivity with protein fragments expressed from deletion mutants in transfected cells. Western blotting, immunoprecipitation and immunofluorescence assays were performed. These MAbs recognized epitopes in different domains of the P protein: 60% were directed against a region lying between residues 83-172 suggesting a major antigenic determinant of the rabies virus P protein in this region. Most of the antigenic sites appeared to be composed of linear epitopes. These MAbs will be useful as tools to dissect structural and functional properties of the rabies virus P protein.

MeSH terms

  • Animals
  • Antibodies, Monoclonal*
  • Antibody Specificity
  • Blotting, Western
  • Cell Line
  • Cloning, Molecular
  • DNA Primers
  • Electrophoresis, Polyacrylamide Gel
  • Epitopes / analysis*
  • Escherichia coli
  • Mice
  • Molecular Chaperones
  • Phosphoproteins / chemistry
  • Phosphoproteins / immunology*
  • Polymerase Chain Reaction
  • Rabies virus / immunology*
  • Recombinant Proteins / chemistry
  • Recombinant Proteins / immunology
  • Viral Structural Proteins / chemistry
  • Viral Structural Proteins / immunology*

Substances

  • Antibodies, Monoclonal
  • DNA Primers
  • Epitopes
  • Molecular Chaperones
  • P phosphoprotein, Rabies virus
  • Phosphoproteins
  • Recombinant Proteins
  • Viral Structural Proteins