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目的 :制备抗p16单克隆抗体并进行鉴定。方法 :以p16合成肽为抗原免疫纯系小鼠。利用杂交瘤技术制备单克隆抗体 ,并利用亲和层析等法纯化抗体。通过酶免疫测定及免疫印迹等法对单克隆抗体进行分析及鉴定 ,并通过免疫组化方法与进口多克隆抗体进行比较。结果 :获得 4株稳定分泌抗 p16单克隆抗体杂交瘤株 ,与进口多克隆抗体比较 ,检测阳性及阴性符合率达 10 0 %。但使用单克隆抗体时 ,组织切片不需进行抗原修复 ,染色效果稳定。结论 :抗p16单克隆抗体的获得弥补了国内 p16检测试剂的空白。
Objective: To prepare anti-p16 monoclonal antibody and identify it. Methods: Pure mice were immunized with p16 synthetic peptide as antigen. Monoclonal antibodies were prepared using hybridoma technology and antibodies were purified by affinity chromatography. Monoclonal antibodies were analyzed and identified by enzyme immunoassay and Western blot, and compared with imported polyclonal antibodies by immunohistochemistry. RESULTS : Four hybridoma strains stably secreting anti-p16 mAbs were obtained. Compared with imported polyclonal antibodies, the positive and negative coincidence rates were up to 100%. However, when monoclonal antibodies are used, tissue sections do not require antigen retrieval and the staining results are stable. Conclusion : The acquisition of anti-p16 monoclonal antibody has made up for the lack of domestic p16 detection reagents.