Preparation Of Monoclonal Antibodies Against An Avian Reovirus Strain Isolated From Semi-Muscovy Duck

Two ARV Strians(ARV-sMd and ARV-Md) isolated from semi-muscovy ducklings and muscovy ducklings respectively were propagated on muscovy duck embryo fibroblast. After Adapted on Pekin duck embryo fibroblast, their serological relationship to chicken arthritis virus and the morphology of ARV-sMd were tested and the results tured out that the virus belonged to avian reovirus. The S3 gene of the two strains were cloned and sequenced and their homology with other ARV strains were analysed.Monoclonal antibodies were prepared against ARV-sMd. Balb/c mice were immunized with purified virus. The immunized splenocyte were collected and fused with SP2/0 myeloma cells.After screening by indirect ELISA and subcloning three times by limiting-dilution, three hybriddoma cell lines named 1D₅, 2D₈ and 3C₃ were established. These hybridomas were able to screte monoclonal antibobies stably, the ELISA titer of the mice ascitic fluid was 1:10⁵ for 1D₅, for 2D₈ and 3C₃ the titers reached 1:10⁷. A McAbs isotyping kit was used to indentify the subclass of these Mabs, results tured out that 1D₅, 2D₈ and 3C₃ belongs IgM, IgG1 and IgG1 respectively.The light chain of these Mabs were categorized into k type. In a western-blot assay with virus proteins, 2D₈ and 3C₃ could react with a protein of molecular weight about 75kd.Further studies on the implication of the Mabs are still needed.