Comparison on Infectious Bursal Disease Monoclona Antibodies Prepared with Two Different Immunogens

摘要

[Objective] To compare the characteristics of monoclonal antibodies against infectious bursal disease virus (IBDV) prepared with two different immunogens,VP2 protein expressed by prokaryotic system and purified IBDV.[Method] IBDV VP2 gene was amplified by RT-PCR and expressed in a prokaryotic system.The recombinant protein was purified by affinity chromatography.IBDV was purified by ultracentrifugation.Balb/c mice were immunized with the purified recombinant protein and IBDV,respectively.The monoclonal antibodies were screened by ELISA.[Result] Two cell lines secreting antibodies against IBDV VP2 protein were obtained,and their ELISA titers were 1:2×104.Four cell lines secreting antibodies against IBDV were produced,and their ELISA titers were 1:2×106,1:6×104,1:1×105 and 1:4×103,respectively.All monoclonal antibodies specifically bound to their own immunogen but did not react with other viruses or proteins.After 10-20 passages,these cell lines still secreted antibodies stably.[Conclusion] The monoclonal antibodies prepared with the recombinant IBDV VP2 protein or purified IBDV can induce immune response in mice,and VP2 specific monoclonal antibodies can be obtained with VP2 protein expressed in the prokaryotic system as immunogen.