Veterinary immunology and immunopathology · 2025
Monoclonal antibodies targeting the Erns protein of classical swine fever virus: application for virus detection.
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Gopinath Shreya, Hosamani Madhusudan, Basagoudanavar Suresh, Sreenivasa B P, Joseph Biljo V, Patil Sharanagouda S
Аннотация
Classical swine fever (CSF), caused by the classical swine fever virus (CSFV; genus Pestivirus), is a highly transmissible disease in pigs that leads to severe economic losses for pig farmers worldwide. Early and accurate diagnosis of CSFV infections is crucial for effective containment of the disease, especially in endemic areas. In this study, the full-length viral protein Erns was successfully expressed in E. coli. The recombinant protein was purified using Ni-NTA affinity chromatography, and its antigenicity confirmed by western blot and indirect ELISA (iELISA) using CSFV-specific antiserum. The purified Erns protein was then used as an immunogen to generate murine hybridomas producing monoclonal antibodies (mAbs). A panel of 15 mAbs targeting this protein was characterized using iELISA and western blotting, for their ability to react with native antigens in cultures infected with CSFV. These mAbs have the potential to be used in the development of diagnostic tools, particularly for the detection and titration of virus-infected cultures through endpoint dilution assays.