Yu A;Chunosheng;Jinfu Yang;Jianchun Jianchun;Guihai Chen;Xiaofeng Xiaofen

mhc ii;monoclonal antibody;000;alpha chain;hybridoma;exon

[Objective] To prepare the monoclonal antibody against chicken major histocompatibility complex class II molecules(MHC II).[Method] The prokaryotic expression of the gene fragments of exons 2-6 encoding alpha chain of MHC II and exons 3-6 encoding beta chain of MHC II were performed based on its protein sequences.After BALB/c mice were immunized with the purified fusion proteins,the mouse spleen cells were fused with mouse myeloma cells SP2/0.Then the positive hybridoma cells were screened and detected by indirect enzyme-linked immunosorbent assay(ELISA).[Result] One hybridoma cell strain secreting monoclonal antibody against alpha chain and two strains secreting monoclonal antibody against beta chain were obtained.These three hybridoma cell strains were named as MHC II alpha-4,MHC II beta6-2 and MCH II beta6-31,respectively.Their titers of ascites in indirect ELISA were 1:256 000,1:256 000 and 1:1 280 000,respectively.These antibodies could specifically recognize MHC II alpha chain or beta chain in western blotting.[Conclusion] Three obtained hybridoma stains can stably produce the monoclonal antibody against chicken MHC class II molecules.