动物造模 z-bio 2022-11-28 446

　　OBJECTIVE: To prepare duck antigen processing-associated transporter (TAP) specific monoclonal antibody, and to provide experimental materials for in-depth use of experimental ducks for immunological research.

　　Methods: The major histocompatibility complex (MHC) haplotype HBW-SPF duck TAP protein peptide-binding region fragment was induced and expressed by Escherichia coli, and the expression product was purified by nickel column and immunized BALB/c mice. Screening of specific monoclonal antibody-secreting hybridoma cell lines. The positive cell lines were prepared in mouse ascites as the primary antibody, and subjected to western blotting test with multiple truncated TAP protein peptide binding regions to identify the antigenic epitopes against which the monoclonal antibody was directed. The reactivity of the monoclonal antibody to the peripheral blood lymphocytes of experimental ducks and chickens was compared by indirect immunofluorescence test, and the specificity of SPF chicken, SPF duck, quail, goose and SPF pig was detected by immunohistochemical technique.

　　RESULTS: A duck TAP monoclonal antibody 1A6 was obtained, with the epitope at 297NARHQMLQQAVLDATAGTGMVVQEAI322, which showed immunofluorescence reactivity to peripheral blood lymphocytes of chickens and ducks; a large number of specific signals were detected in the intestinal mucosa propria of chickens and ducks; No signal was detected in quail and geese.

　　Conclusion: A monoclonal antibody specific for the antigen transport-related body with chicken and duck reactivity has been obtained, which can be used in the research of avian laboratory animals in avian disease and avian immunology.