抗人膀胱癌抗独特型抗体的制备和鉴定

Production and identification of anti-idiotypic antibody of anti-human bladder carcinoma

目的 制备抗人膀胱癌抗独特型抗体 (Ab2 )并进行体外实验和鉴定 ,探讨通过免疫效应治疗或预防膀胱癌术后复发的可能性。方法 提取膀胱癌抗原 (Ag) ,应用膀胱癌抗原通过杂交瘤技术制备抗人膀胱癌单克隆抗体 (Ab1 )。 Ab1 经胃蛋白酶水解制备 Ab1 的 F(ab') 2片段 ,以 F(ab') 2片段免疫新西兰白兔 ,其血清经凝胶纯化后获抗人膀胱癌抗独特型抗体 (Ab2 )。采用 ELISA等方法进行鉴别。结果 Ab2 与 Ab1 呈阳性反应 ,与 BALB/ C小鼠γ球蛋白呈阴性反应 ;Ab2 与 Ag竞争结合 Ab1 ;Ab2 抑制 Ab1 与 Ag的结合。结论 Ab1 是针对膀胱移行细胞癌的单克隆抗体 ;Ab2是具有膀胱癌抗原内影像的抗独特型抗体。

Objective To produce anti idiotypic antibody of anti human bladder carcinoma and to identify it Methods Bladder carcinoma antigen (Ag) was drawn from human bladder transitional cell carcinoma by use of even grind and gel Monoclonal antibody of anti human bladder carcinoma (Ab 1) was produced by the use of the bladder carcinoma antigen through B lymphocyte hybridoma technology F(ab`)2 fragment from Ab 1 was prepared through pepsin cleavage Anti idiotypic antibody of anti human bladder carcinoma (Ab 2) was produced by the F(ab`)2 fragment from Ab 1 immuning rabbits Its serum was purified through gel Then it was identified through ELISA Results The reaction of Ab 2 was positive with Ab 1,while it was negative with murine gamma globulin through indirecte ELISA test Ab 2 competitively bind Ab 1 with Ag Ab 2 competitively inhibited the binding of Ab 1 with Ag Conclusions Ab 1 was monoclonal antibody of human bladder transtional cell carcinoma Ab 2 was an anti idiotypic antibody bearing internal image of human bladder antigen