Development Of The Monoclonal Antibodies Against Hemagglutinin Of H3 And H11 Subtype Avain Influenza Virus

Avian influenza (AI) is a contagious disease of several animals including humans,pigs and poultry. H3 and H11 subtype AIV has a high isolate rate in ducks, and H3 is the main subtype of human influenza virus. At present, the hemagglutination inhibition(HI) test, a convenient method, was widely used in the diagnosis of AIV infection. In this study, the monoclonal antibodies against hemagglutinin(HA) of H3 and H11 subtype AIV were developed for the HI test.The eight-week-old female BALb/c mice were immunized with H3 and H11 subtype AIV. Spleen cells were collected and fused with Sp2/0-Ag-14 myeloma cells after the third immunization. Hemagglutinin inhibition (HI) test was used to screen hybridoma cells and limited dilution method was performed to subclone the positive clones. After two cycles of subcloning, four McAbs against the HA of H3-AIV were obtained, and were designated as 1D7,1F9,5A4 and 5F5, respectively. One McAb against the HA of H11-AIV was obtained, and was designated as 3F10. They were proved to have hign HI titers and were specific to AIV-H3 and AIV-H11, but not reacted with other subtypes of AIVs and NDV. We chose 28 H3-AIV isolates to react with the four McAbs by the HI test. 1F9,5A4,5F5 reacted with all the 28 virus strains, but 1D7 only reacted with 26 of the 28 virus strains. We chose six H11-AIV isolates to react with 3F10 by the HI test. It reacted with all the six virus strains. These results showed that these McAbs could replace the chicken serum in epidemiological investigation of H3-AIV and H11-AIV, and will be useful in controlling influenza of both animals and...