Development of the monoclonal antibodies against hemagglutinin of H3 subtype avian influenza virus

The female mice of eight-week-old were immunized with subtype H3-AIV. Spleen cells were collected and fused with SP2/0 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 hemagglutinin of H3-AIV were obtained, and designated as 1D7,1F9,5A4 and 5F5, respectively. They were proved to have high HI titers and were specific to AIV-H3, but not to react with other subtypes of AIVs and NDV. We chose 28 H3-AIV isolates to react with the four McAbs by the HI test. The McAbs of 1F9,5A4 and 5F5 reacted with all the 28 virus strains, but 1D7 only reacted with 26 of 28. It suggested that the four McAbs can be used in epidemiological investigation of H3-AIV, and will be useful in controlling influenza of both animals and human beings.