鼠抗人B7-H4单克隆抗体的制备及功能研究  

The Preparation of Anti-Human B7-H4 Monoclonal Antibody and Its Function Study

作　　者：张标[1] 房鹏[1] 张宁[1] 顾宗江[1] 

机构地区：[1]苏州大学医学部免疫学系,江苏苏州215007

出　　处：《中国血液流变学杂志》2015年第1期18-21,共4页Chinese Journal of Hemorheology

摘　　要：目的：制备鼠抗人B7-H4单克隆抗体（mAb），并对其生物学功能进行初步研究。方法以表达B7-H4分子的转基因细胞L929/B7-H4为免疫原，免疫Balb/c小鼠，采用杂交瘤技术，融合小鼠骨髓瘤细胞与脾脏细胞，用流式细胞术筛选阳性克隆，获得稳定分泌抗B7-H4 mAb的杂交瘤细胞株，并对mAb的生物学功能进行初步研究。结果成功获得1株稳定分泌抗人B7-H4 mAb的杂交瘤细胞株，命名为8F10。Western blot结果显示：8F10特异性识别B7-H4蛋白；T细胞增殖抑制阻断实验显示8F10可以阻断B7-H4对T细胞增殖的抑制作用。结论成功获得鼠抗人B7-H4 mAb，为进一步研究B7-H4分子的生物学功能提供了有效的工具。ObjectiveTo prepare anti-human B7-H4 monoclonal antibody （mAb） and study its biological functions.Methods The L929/B7-H4 transfectants were used as immunogen to immunize Balb/c mice. After the fusion of murine myeloma cells with spleen B cells of the mice through hybrid technology, FCM was employed to screen positive clones and to obtain hybridoma cell line stably secreting mAb against human B7-H4, of which biological functions were studied.Results One hybridoma cell line stably secreting specific anti-human B7-H4 was established, named 8F10. Western blot result showed that 8F10 could specifi cally bind to human B7-H4 protein. The results of blocking T cell proliferation inhibition displayed that mAb 8F10 could block the inhibitory effects of B7-H4 on T cell proliferation.Conclusion One hybridoma cell line secreting anti-B7-H4 mAb was obtained. This mAb provides useful tool for further studies of B7-H4 biological function.

关 键 词：B7-H4 单克隆抗体 杂交瘤 B7-H4 

分 类 号：Q813.2[生物学—生物工程]