Preparation and Characterization of Monoclonal Antibody Specific to Hemagglutinin of Subtype H5 Avian Influenza Virus

Objective To develop and characterize monoclonal antibody (McAb) against AIVH5 for related research work.Methods BALB/c mice were immunized conventionally with the purified AIVH5 On the third day after the final immunization, spleen cells of mice fused with myeloma cells SP2/0 were filtered by culturing selectively, diluting finitely and hemagglutination_inhibition (HI) test. The subtypes were obtained from the clone line by ELISA test and their characteristics and mantles were determined by H5, H7 and H9 subtypes of 37 AIV lines. Results The three cell lines secreting McAbs stably by culturing in vitro and recovery tests were obtained and named as 1E 5 , 4A 4 , and 4B 1 respectively. The subtypes of the three McAbs were identified as IgG1, Kappa light chain. The HI titers of culture supernatants and ascites were 1∶2 6 -1∶2 8 and 1∶2 10 -1∶2 16 respectively. The three McAbs reacted specifically with only AIVH5 but not with AIVH9, AIVH7, NDV in chickens, NDV in geese, EDS76 and so on. Conclusion The three McAbs will be very useful as specific reagents in diagnosing and controlling avian influenza.