[Preparation and characterization of monoclonal antibodies against M2 protein of avian influenza virus].

AIM

To prepare and characterize the monoclonal antibodies against M2 protein of avian influenza virus (AIV).

METHODS

BALB/c mice were immunized with purified fusion protein GST-M2. Then the hybridoma cell lines secreting mAb against M2 were screened by indirect ELISA by using GST-M2 and GST as ELISA coated antigen respectively and selecting strongly positive against GST-M2 and negative against GST.

RESULTS

Four hybridoma cell lines secreting anti-AIV M2 mAbs were obtained, designated 1E1, 2F8, 4E3 and 5D6. The ELISA titers of these mAbs were above 2(10) x 100 and the agar gel precipitation test showed the titers of them were above 1:4. Immunoglobulin subclass tests differentiated them as IgG2a (1E1, 4E3) and IgG2b (2F8, 5D6). The Ag-Capture ELISA showed the mAb 2F8 could specifically reacted with H5 and H9 subtypes of AIV, but could not react with Newcastle disease virus (NDV) and Infectious bursal disease virus (IBDV). IFA and immunochemistry test indicated the prepared mAb 2F8 could specifically bind to MDCK cell and chicken tissue cells infected with different AIV subtypes.

CONCLUSION

Four mAbs against AlV M2 were obtained in this study. The mAb 2F8 against AIV M2 could be used as the key reagent for establishment of AIV detection method.