人CTLA4单克隆抗体的制备和鉴定  

Preparation and identification of monoclonal antibody against human CTLA4

作　　者：卢洋[1] 钱卫珠[1] 王皓[1] 吴孟超[1] 郭亚军[1] 

机构地区：[1]第二军医大学肿瘤研究所东方肝胆外科研究所,上海200433

出　　处：《医学研究生学报》2002年第2期98-101,共4页Journal of Medical Postgraduates

基　　金：国家杰出青年自然科学基金 ( 3992 5 0 2 1) ;上海市现代生物与新药产业发展基金 ( 9943192 0 4)

摘　　要：目的 :制备人的细胞毒性T淋巴细胞相关抗原 4 (CTLA4 )的单克隆抗体。　方法 :利用蛋白纯化技术获得人CTLA4Ig蛋白免疫Balb/c小鼠。取免疫小鼠的脾细胞与骨髓瘤细胞NS1融合 ,用ELISA法筛选分泌单克隆抗体的杂交瘤细胞株 ,并用Western印迹和流式细胞检测仪鉴定所制备的单克隆抗体。　结果 :获得一个稳定的分泌抗人CTLA4的杂交瘤细胞株。该细胞克隆培养上清中所含的单抗能和经过PHA刺激后的人T细胞结合 ,但不能和经过PHA刺激后的小鼠T细胞结合。　结论 :利用重组人CTLA4Ig制备获得高特异性的抗人CTLA4单克隆抗体。Objectives:To prepare monoclonal antibody specific for human CTLA4 molecule. Methods: Balb/c mice were immunized with purified human CTLA4 Ig. The spleen cells from immunized mice were fused with mouse myeloma cell line NS1. Resulting hybridomas were screened for production of anti huCTLA4 antibodies by ELISA procedure. The obtained monoclonal anti huCTLA4 antibodies were further analyzed with western blot and flow cytometry. Results:One hybridoma clone stably secreting anti huCTLA4 antibodies was obtained. The monoclonal antibodies in the cell culture supernatant can specifically bind to the PHA stimulated human T cells, while cannot bind to the PHA stimulated murine T cells. Conclusions:Monoclonal anti huCTLA4 antibody was obtained using this immunization procedure with huCTLA4 Ig.

关 键 词：CTLA4 单克隆抗体 T细胞免疫 制备 鉴定 

分 类 号：R392-33[医药卫生—免疫学] R446.62[医药卫生—基础医学]