Preparation And Of MHC Class? Monoclonal Antibody Specific To B19

The major histocompatibility complex(MHC)of chickens refers to a group of closely linked,highly polymorphic genomes on chicken chromosome 16.Almost all nucleated cell surfaces express major histocompatibility complex class I molecules(MHC I).MHC class I molecules are dimers that are non-covalently bound by transmembrane glycoprotein heavy chain alpha and light chain ?2m microglobulin.The MHC class I gene(BF gene)is responsible for encoding the heavy chain alpha,and the molecular weight of the mature alpha chain is about 44 kDa.In this study,6-week-old BALB/c mice were immunized with prokaryotic and purified BF protein.After 72 hours of immunization,spleen cells were fused with SP2/0 cells.Hybridoma cells were screened by indirect ELISA.After 2-3 subclones,And two high-specific BF monoclonal antibodies were named as 1F12 and 3C10 respectively.The heavy chain was IgM and the light chain was ? chain.1F12 and 3C10 hybridoma cells were inoculated with 8-week-old BALB/C mice,to prepare ascites.The results of Western Blot showed that the prepared monoclonal antibody reacted only with the corresponding recombinant protein and did not react with the uninduced bacteria,and showed good specificity.The epitope of 1F12was identified by truncated expression in P66-Q83.The monoclonal antibodies provide useful experimental basis for further study on the function of BF in immunologyThe BW/G1,BW/G2,BW/G3,BW/G6 and BW/G7 strains of MHC haploid chickens cultivated and preserved by the National Veterinary Research Animal Seed Center of the Chinese Academy of Agricultural Sciences,Harbin Institute of Veterinary Medicine,Chinese Academy of Agricultural Sciences,including MHC B5,B15,B2,B21 and B19.According to the BNK gene sequence of different haplotype chickens published on NCBI,the BNK sequences of different haplotypes were compared.The results of biological analysis of different haplotypes were consistent with the standard haplotype.The results showed that BNK gene provided the experimental basis for the monitoring of genetic stability of MHC haplotype chickens.This study provided the scientific basis for establishing the importance of MHC-specific haplotype SPF flocks,and laid a foundation for understanding the relationship between chicken MHC genetic variation and disease resistance.At the same time,it provides a basis for further study on the biological function of different haplotypes.