Development, characterization and application of monoclonal antibodies against woodchuck hepatitis virus core protein

Objective:To develop monoclonal antibodies against woodchuck hepatitis virus core protein which can be used to detect woodchuck hepatitis virus, and to carry out research on woodchuck hepatitis virus and related hepatitis virus.Methods:BALB/c mice were immunized with prokaryotically expressed recombinant woodchuck hepatitis virus core protein(WHc 1-149), mouse spleen cells were fused with SP2/0 cells by a standard hybridoma technique. Positive hybridomas that produced antobodies against woodchuck hepatitis virus core protein were screened by ELISA and immunohistochemistry. Cloning procedure was carried out using the method of limiting dilution.Results:5 hybridoma clones stably producing monoclonal antobodies(4B1E,6C5D,6C5C,6D1D,6D1G) against woodchuck hepatitis virus core protein were established. All these antibodies were shown to be good for ELISA, IHC, and Western blot assays and found cross-reactive between HBcAg and 5 monoclonal antobodies by our experiments. Using these antibodies we were able to detect positive signal in liver tissue of some Chinese Marmots by IHC.Conclusion:All these 5 monoclonal antibodies we developed can be used for WHV or Hepadnavirus study, and may also serve as an important tool for discovering new relative hepadnaviruses.