Production and characterization of monoclonal antibodies specific for pseudorabies virus

Abstract

Monoclonal antibodies (MAbs) against Pseudorabies virus (PrV) were obtained by the fusion of P3x-Ag8.653 myeloma and spleen cells from immunized BALB/c mice with a suspension of Pseudorabies (PrV) virus strains: MAVE (Morbus Aujeszk'y virus Ercegovac) and NS 257 (Novosadski virus strain). A total of 95 antibody-secreting hybridoma cells against the virus strain (MAVE and NS 257) of Pseudorabies virus have been isolated. Ten of these monoclonal antibodies were found by ELISA (Enzyme-linked immunosorbent assay) to react specifically with both virus strains. MAbs for VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 were purified by chromatography on protein G Sepharose 4FF and they have been shown to have a strong reactivity in the ELISA test. All MAbs were characterized by electrophoresis SDS-PAGE and electrophoresis Western-blot immunoassay. MAbs VAM 2.1, VAM 4.1, VAM 5.1 and VAM 6.1 in hybridoma culture supernatants and ascites fluid were quantified using a dot-blot immunobinding assay. The VAM 2.1 MAb was found to be more specific in the reaction with viruses of both strains. The glycoprotein of 40 kD molecular weight was found on the surface of virus strain MAVE. Results showed that the produced and characterized MAbs against PrV strains can be used for the detection of Aujeszky's disease.