Generation of Monoclonal Antibody to Avian Influenza Virus

The BALB/c mice were immunized four times with the inactive purified subtype H5 avian influenza virus(AIV-H5),then the spleen cells of the hyperimmunized mouse and SP2/0 cells were fused with PEG-1500.The cell fusion rate of 65.8% was obtained.An indirect enzyme-linked immunosorbent assay(ELISA) was developed to detect monoclonal antibodies(McAbs) secreted by hybridoma cell lines. And a positive rate of 14.2% was obtained.Two hybridoma cell lines secreting the monoclonal antibodies to AIV,named 5A6,5G7 were developed by two subclones.The specificity of McAb was identified by means of indirect ELISA.After repeated frozen storage、thawed revival and generation,they were still steadily producing high titer of McAbs.The results of latex agglutination test showed that the antibodies secreted by 5A6 and 5G7 cells belong to IgG1,κisotype.The McAbs prepared would be very useful in diagnosis and control of avian influenza.