Preparation and Identification of Monoclonal Antibodies Against Haemagglutinin of Subtype H5 Avian influenza virus

Balb/c mice were immunized with the purified AIV antigens.Splenocytes from the immunized mice were fused with SP2/0 myeloma cells.Positive hybridoma clones were screened by hemagglutination inhibition(HI),and limited dilution method was performed to subclone the positive clones.After three cycles of subcloning,seven McAbs against the hemagglutinin of H5-AIV were obtained,designated as 1C4,1D4,1E12,2E11,4C12,4G2 and 5E12.The HI titers of culture supernatant and ascites were 2~(4)-2~(7) and 2~(10)-2~(18).The seven McAbs had good specificity to H5 subtype of avian influenza virus (H5-AIV).No cross-reactions were found when H7 amd H9 subtypes of avian influenza virus,Newcastle disease virus(NDV) and Avian infectious bronchitis virus(IBV) were tested.All McAbs had high neutralization titers.