Development And Characterization Of Monoclonal Antibodies Against H5, H7 Hemagglutinin And Nucleoprotein Of Avian Influenza Virus

Highly pathogenic avian influenza (HPAI) is a devastating disease of poultry leading to high mortality caused by some H5 and H7 subtypes influenza A viruses. HAPI is classified as one of the list A animal disease by OIE and China Ministry of Agriculture. As it has a great economic impact on the poultry industry and it also can cause severe human disease, a rapid and effective detection assay is urgently required. In this study, hybridoma cell lines secreting mAbs against nucleoprotein of avian influenza viruses and hemagglutinin protein of H5 and H7 subtypes of avian influenza viruses were generated. These mAbs would be very useful for the rapid detection of avian influenza virus.To prepare the mAbs against the hemagglutinin protein of H5 or H7 subtype avian influenza virus (AIV) and the mAbs against the nucleoprotein of avian influenza virus, 6～8 weeks old BALB/c mice were immunized endermicly with H5 or H7 subtype AIV or immunized intramuscularly with purified recombinant plasmid pcDNA3.1-NP. Splenocytes from the immunized mice were fused with Sp2/0-Ag-14 myeloma cells after the fourth immunization, hemagglutinin (HA), hemagglutinin inhibition (HI) test and indirect ELISA was used to screen hybirdoma cells. Nine hybirdoma cell lines specific antibodies against the HA protein of H5 subtype AIV were established, four hybirdoma cell lines for the HA protein of H7 subtype AIV, and five hybirdoma cell lines for nucleoprotein of AIV were also developed.