Preparation Of Monoclonal Antibodies Against The H5 Haemagglutinin Of Avian Influenza Virus

H5 avian influenza virus was cultured in chicken embryo. The obtained virus was inactivated with 0.1% formaldehyde and then purified by the methods of differential centrifugation and running Sephadex-G 200 . Balb/c mice were immuned with the purified AIV antigens. Splenocytes of vaccinated Balb/c mice were fused with the mouse SP2/0 myeloma cells. The hybridrate was 99.1%. Two hybridomas 3D7 and 2G4 secreting monoclonal antibody(McAb) against H5 haemagglutinin of avain influenza virus were obtained after screened by haemagglutination ihibition test (HI) and cloned three times by limiting dilution.Analysis with colchicine indicated that the number of chromosome in the two hybridomas 3D7 and 2G4 cells were about 98 and 96 respectively. Both the two McAbs were proved to belong to immunoglobulin subclass IgG1. The HI titers of cell culture fluid of 2G4 and 3D7were 1:2⁷ andl:2⁹ and those of ascites were l:2¹¹and 1:2¹² respectively. Both of the two monoclonal antibodies were able to secret McAb stably and they were only bind to H5, no crossreaction with NDV, H7, H9 and EDSV. Both two McAbs have good neutralization activity.After twenty-five serial generations or resuscitated after 3 months of deepfreeze, the two hybridomas were still able to secret monoclonal antibody (McAb) against H5 haemagglutinin of avain influenza virus stably, as indicates that the two hybridomas have good stability.