抑制胃癌增殖功能性单抗的制备、筛选及鉴定  

Preparation,screening and identification of inhibition proliferation functional monoclonal antibodies of gastric cancer

作　　者：赵西路[1] 陈立钊[1] 冉宇靓[1] 胡海[1] 孙立新[1] 遇珑[1] 杨治华[1] 

机构地区：[1]中国医学科学院北京协和医学院肿瘤研究所细胞及分子生物学研究室,北京100021

出　　处：《中国卫生检验杂志》2008年第6期1013-1015,共3页Chinese Journal of Health Laboratory Technology

基　　金：国家自然科学基金面上资助项目(30570818);国家重点基础研究发展规划(973)项目(2002CB513106)资助

摘　　要：目的:批量制备、筛选、鉴定抑制人胃癌细胞增殖的功能性单克隆抗体,为筛选获得与胃癌增殖相关的基因/蛋白奠定基础。方法:以人胃癌细胞系NCI-N87免疫BALB/c小鼠,经脾细胞融合后制备单抗,活细胞免疫荧光、免疫组化、细胞增殖实验、ELISA、Western blotting等方法筛选鉴定抑制胃癌生长的功能性单抗。结果:共获得1012株杂交瘤,252株能与胃癌细胞膜反应,145株与正常胃组织不反应或低反应。11株单抗在体外能够显著抑制N87的增殖。其中1株单抗1B7属IgMκ类,其识别的抗原的相对分子质量为32kDa。结论:成功获得了多株能够抑制胃癌细胞增殖的功能性单抗,其中1株单抗识别的抗原可能具有作为胃癌靶向治疗靶标的潜在应用价值。Objective：Preparation, screening and identification of monoclonal antibodies which can inhibit gastric cancer cells proliferation may provide a foundation for separation and acquirement of genes or proteins related to gastric cancer growth. Methods： Gastric tumor cells NCI - N87 were used to immunize BALB/c mice. The spleen cells of the immunized mouse were fused with SP2/0 cells. So as to obtain functional mAbs, antibodies of each clone were screened by immunofluorescence, immunohistochemistry, tumor proliferation assay, ELISA and western blotting. Results：A total of 1012 clones of mAb were obtained by the fuse of spleen cells with SP2/0 cells. 252 clones of mAb can interact with the membrane of the gastric carcinoma cells N87 by viable immunofluorescence. 145 clones weekly or barely reacted with normal gastric tissue. 11 of these antibodies could significantly inhibit gastric cancer cells proliferation in vitro. One of these antibodies 1B7, which was identified as IgMK, could noticeably suppress N87 cells proliferation in vitro. Western blotting displayed that this clone reacted with antigen of 32kDa. Conclusion： We successfully obtained several functional mAbs which suppressed the proliferation of gastric carcinoma cells in vitro. The antigen recognized by a functional mAb may be a potential target of targeted therapy of gastric carcinoma.

关 键 词：胃癌 功能性单抗 增殖 靶向治疗 

分 类 号：R730.5[医药卫生—肿瘤]