抗主要组织相容性相关蛋白MICA/B的单克隆抗体制备及应用

Production and application of monoclonal antibody against major histocompatibility complex class Ⅰ related chain A and B

目的:制备抗主要组织相容性相关蛋白A/B(MICA/B)的单克隆抗体并对其特异性进行验证。方法:以重组MICA*012蛋白免疫小鼠后将其脾细胞与SP2/0细胞进行融合和筛选得杂交瘤细胞株,并用ELISA对其中9B10细胞株产生的单克隆抗体进行效价检测以及通过Western blot、Luminex、免疫荧光对其特异性进行检测。结果:获得6株杂交瘤细胞株,其中9B10细胞株产生单克隆抗体(m Ab)9B10的最低反应浓度为0.02 ng/μl,且m Ab 9B10可应用于Western blot、Luminex和免疫组化荧光试验对MICA和MICB分子的检测。结论:成功获得可同时检测MICA和MICB表达的单克隆抗体9B10。

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 r MICA＊ 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 m Ab 9B10 was 0. 02 ng/μl,and the specificity of m Ab 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.