Production and application of monoclonal antibody against major histocompati-bility complex class Ⅰrelated chain A and B / 中国免疫学杂志

ABSTRACT

Objective:Generation and identification of hybridoma to produce monoclonal antibody against MICA/B. Methods:SP2/0 cells were fused with murine splenocyte immunized with recombinant protein rMICA?012 to get hybridoma cell lines. The titer of the monoclonal antibody produced by 9B10 cell line was determined by ELISA and its specificity was tested by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence assay. Results:Six hybridoma cell lines were selected by ELISA screening test. The minimum reaction concentration of mAb 9B10 was 0. 02 ng/μl,and the specificity of mAb 9B10 was determinated by Western blot,Luminex mutiplex microsphere-based immunoassay and immunofluorescence. Conclusion:The monoclonal antibody 9B10 was available to apply for the detection of MICA and MICB expression.