[Preparation and identification of monoclonal antibodies against recombinant N protein of mouse hepatitis virus].

AIM

To prepare and characterize the monoclonal antibodies (mAbs) against N protein of mouse hepatitis virus (MHV).

METHODS

BALB/c mice were immunostimulated with the purified recombinant N protein expressed by Bac-to-Bac baculovirus expression system. Then the spleen cells of the immunized mice were fused with Sp2/0 myeloma cells by conventional hybridoma techniques. An indirect ELISA using recombinant N protein as antigen was established to screen antibody-producing hybridoma cell lines. Western blotting and IFA were applied to characterize the mAbs.

RESULTS

We obtained four hybridoma cell lines secreting mAbs against recombinant N protein. The subclasses of two mAbs were IgG2a, one IgG2b and the other IgG1, and light chains were both Kappa. Western blotting showed that the mAbs specifically recognized certain antigenic epitope of N protein of MHV.

CONCLUSION

mAbs against N protein of MHV with high activity and specificity were prepared successfully, which provides a potential basis for further researches on N protein function and diagnostic methods.