抗人膀胱癌双功能抗体的制备与鉴定

目的:制备抗人膀胱癌双功能抗体(BfAb)并进行体外实验与鉴定.方法:提取膀胱癌抗原,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,后者经胃蛋白酶水解后制备抗人膀胱癌单克隆抗体的F(ab′)2片段.然后将该片段通过二次杂交制备杂交瘤,用其免疫新西兰白兔,血清经凝胶纯化后获抗人膀胱癌BfAb,应用ELISA和免疫组化方法进行鉴别.结果:抗人膀胱癌BfAb与抗人膀胱癌单克隆抗体呈阳性反应,与BALB/C小鼠γ球蛋白呈阴性反应;BfAb可以同时结合靶细胞的肿瘤抗原和效应细胞表面的触发分子,引发正常细胞对肿瘤细胞的防御机制.结论:BfAb既能竞争性地与膀胱移行细胞癌抗原结合,又具有激活淋巴细胞的功能,从而特异性的直接杀伤癌细胞并诱发一系列的免疫应答反应.

Preparation and identification of bifunctional antibodies against human bladder carcinoma

Objective:To prepare and identify bifunctional antibody (BfAb) against human bladder carcinoma. Methods:Bladder carcinoma antigen (Ag) was prepared from human bladder transitional cell carcinoma by use of even grind and gel. Monoclonal antibody of anti human bladder carcinoma (Ab 1) was prepared by the use of the bladder carcinoma antigen through B lymphocyte hybridoma technique. F(ab′) 2 fragment from Ab 1 was prepared through pepsin cleavage.Rabbits were immunized with the second hybridoma of Ab 1 and F(ab′) 2 fragment. Antiserum, the BfAb against human bladder cancer, was purified through gel and then was identified through ELISA and immunohistochemistry. Results: The reaction of BfAb with Ab 1 was positive, while it was negative with murine gamma globulin .BfAb could bind with Ag of target cells and trigger molecules at the same time.Normal protective mechanism against cancer cells were thus induced. Conclusion:BfAb could competitively bind with Ag and stimulate lymphocytes.It could directly destroy cancer cells and induce a series of immune response.