Aim: To prepare the monoclonal antibodies (mAbs) specific for nuclear protein (NP) of avian influenza virus (AIV) and identify their biological properties.
Methods: BALB/c mice were immunized with AIV (formaldehyde-inactivated AIV H9N2, Triton X-100-lysed H9N2 and AIV NP expressed in E.coli, respectively). Hybridoma cell lines secreting anti-AIV NP mAbs were developed through cell fusion, screening and cloning. The mAb's titer was determined by indirect ELISA. Specificity of mAbs was identified by cross-reaction test and indirect immuno-fluorescence assay (IFA).
Results: 6 hybridoma cell lines secreting anti-AIV NP mAbs were obtained, designated 4F4, 1C3, 1G11, 1C2, 1D10 and 2F7. ELISA detection showed that the titers of two mAbs (1G11 and 1D10) out of 6 mAbs were the highest (2(-13) and 2(-14), respectively) and their specificity was also better than that of the others, confirmed by cross-reaction test and IFA.
Conclusion: In this study 6 mAbs against AIV NP were obtained. The mAbs 1G11 and 1D10 perform the best in titer and specificity. This work paves the way for AIV study and development of method for rapid detection of AIV.