抗p16单克隆抗体的制备及鉴定

Preparation and Characterization of Monoclonal Antibodies against p16

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 the monoclonal antibodies (McAbs) against p16 and to characterized the McAbs. Methods: Balb/C mice were immunized with polypeptide fragment of p16 as antigen. Hybridoma technique was used to prepare McAbs agaist p16, and affinity chromatography was used to purify the McAbs. The McAbs were analysed and characterized with EIA and Western Blot et al. and were compared with imported polyclonal antibodies (PoAbs). Results: Four hybridoma cell lines which stably secrete McAbs against p16 were got. There was a complete correspondence between the McAbs and imported PoAbs. Conclusion: The McAbs have high specificity and sensitivity. In addition, the prepared monoclonal antibodies against p16 make up the blank space of domestic test reagent.