鼠抗人B7-H4单克隆抗体的制备及初步鉴定  

Preparation and primary identification of mouse anti-human B7-H4 monoclonal antibody

作　　者：房鹏[1] 徐慧[1] 刘艳平[1] 邵联波[1] 侯爵[1] 顾宗江[1] 

机构地区：[1]苏州大学生物技术研究所,江苏省215007

出　　处：《江苏医药》2012年第12期1386-1388,共3页Jiangsu Medical Journal

基　　金：江苏省肿瘤免疫转化医学中心(XK201135)

摘　　要：目的制备鼠抗人B7-H4单克隆抗体,并对其生物学特性进行初步鉴定。方法用稳定表达B7-H4分子的L929/B7-H4转基因细胞免疫6-8周龄的Balb/c小鼠。取脾脏细胞与小鼠骨髓瘤细胞融合后,采用流式细胞术筛选阳性克隆,制备稳定分泌抗B7-H4单克隆抗体的细胞株,并用Western blot和Dot blot鉴定其生物学特性。细胞计数试剂盒8(CCK-8)测定T细胞增殖情况。结果成功获得1株稳定分泌抗人B7-H4单克隆抗体的杂交瘤细胞株,命名为5C8。Western blot和Dot blot结果表明,5C8能与人B7-H4蛋白特异性结合。CCK-8实验显示,5C8可以阻断B7-H4分子对T细胞增殖的抑制作用。结论成功获得鼠抗人B7-H4单克隆抗体,为研究B7-H4分子在肿瘤中的作用提供了手段。Objective To prepare mouse anti-human B7 H4 monoclonal antibody（mAb） and primarily characterize its biological functions. Methods The L929/B7-H4 transgenie cells stably expressing B7-H4 molecule were used to immunize Balb/c mice. After the spleen B cells in the mice were fused with routine myeloma cells, the positive clones were screened by flow cytometry. The cell line secreting anti-B7 H4 mAb was prepared and its biological function was identified by Western blot and Dot blot. The T cell proliferation was detected by cell counting kit-8 （CCK8）. Results One hybridoma cell line steadily secreting specific anti-human B7-H4 was successfully obtained, which was named 5（28. Western blot and Dot blot showed that 5C8 could specifically bind to human B7-H4 protein. CCK8 experiment revealed that the mAb 5C8 could block the inhibitory effect of B7-H4 on T cell proliferation. Conclusion The mouse anti-human B7-H4 mAb is generated and provides an useful tool for further studying： biological function of B7-H4 in canoor

关 键 词：B7-H4单克隆抗体 杂交瘤 

分 类 号：Q813[生物学—生物工程]