Development And Identification Of Monoclonal Antibodies Against H5N1 Subtype Avian Influenza Virus

Avian influenza is a group of symptoms caused by influenza virus A. Avian influenza has raised great concerns for its economic importance to poultry industry and potential threat to the public health. In this experiment, a highly pathogenic avian influenza virus (H5N1), was chosen to immunize Balb/c mice in order to product the monoclonal antibodies (McAbs).The 8-week-old female Balb/c mice were immunized with the allantoic fluid, which was purified and concentrated by differential centrifugation. The spleen cells of immunized mice were fused with SP2/0 myeloma cells. Indirect enzyme linked immunosorbent assay (ELISA) was used to screen hybridoma cells and limited dilution method was performed to subclone positive clones.Five of McAbs specifically against avian influenza virus (AIV) were obtained, designated as 2C1, 4F10, 5A5, 5C12 and 6F5 respectively. One of these 5 McAbs, 4F10, was of IgM subtype. The others were of IgG1 subtype. The titers of McAbs substrates were: 1:640,1:640,1:1 280,1:1 280,1:2 560. The titers of ascites were: 1:12 800,1:160,1:51 200,1:128 000,1:256 000. The result of chromosome analyse was that the average chromosome number of hybridoma cells were 98 to 108.The results of the cross-reaction test suggest that these McAbs were specific to AIV-H5N1 without detectable cross-reactivity with specific pathogen free (SPF) chick embryo allantoid fluid, AIV-H9, Newcasttle disease virus, Eggs drop syndrome virus and Avian infectious bronchitis virus. The results of superposable test suggest that 5A5, 5C12 and 6F5 were against different antigen epitope.These McAbs against AIV-H5N1 will be very useful in control of avian influenza.