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以我室用基因工程表达的人CTLA 4蛋白[1,2 ] 作为免疫原 ,采用细胞融合、ELISA法和免疫印迹等方法制备并筛选到鼠抗人CTLA 4的单克隆抗体 ,并以单向混合淋巴细胞刺激反应 (MLR )测定了抗人CTLA 4抗体对淋巴细胞增殖的影响。结果表明我们得到了 4株单克隆抗体 (1H7、 2G2、 2F11、 3A7) ,它们均可与近似天然构象的人CTLA 4Ig融合蛋白结合 ,又可与大肠杆菌表达出的经复性后的人CTLA 4蛋白结合 ,并且可分别识别人CTLA 4Ig融合蛋白不同抗原决定簇。其中一株(3A7)既和近似天然构象的蛋白结合又可与变性的蛋白结合 ,并具有显著阻断CTLA 4的抑制性免疫信号传递 ,促进同种异体混合淋巴细胞增殖的功能
Using human CTLA4 protein [1,2] genetically engineered in our laboratory as immunogen, the monoclonal anti-human CTLA4 monoclonal antibody was prepared and screened by the method of cell fusion, ELISA and western blot. Mixed lymphocyte stimulating response (MLR) was used to determine the effect of anti-human CTLA4 antibody on lymphocyte proliferation. The results showed that we obtained four monoclonal antibodies (1H7, 2G2, 2F11, 3A7), which can be combined with the approximately native conformation human CTLA 4Ig fusion protein, but also E. coli expression of refolded human CTLA 4 protein and recognize different epitopes of the human CTLA4Ig fusion protein, respectively. One of them (3A7) binds both to the natively conformational protein and to the denatured protein and has the ability to significantly block the CTLA 4 inhibitory immune signaling and promote the proliferation of allogeneic mixed lymphocytes