应用木瓜酶消化抗甲胎蛋白单克隆抗体制备F(ab′)_2片断  

Preparation of F(ab')_2 Fragments by Digesting Anti-AFP McAb with Papain

作　　者：于鲁东 蒋先敏[1] 程明[1] 

机构地区：[1]卫生部北京生物制品研究所,北京100024

出　　处：《中国生物制品学杂志》1994年第4期148-150,共3页Chinese Journal of Biologicals

摘　　要：用胃酶消化抗甲胎蛋白单克隆抗体（简称AFPMcAb）制备F（ab′）2片断，反应时间长，且F（ab′）2产量低。改用半胱氨酸活化的木瓜酶，在没有半胱氨酸存在的条件下消化AFPMcAb。反应4小时后．SDS－PAGE电泳结果显示，90％以上的AFPMcAb被降解为F（ab′）2片断、经S－200层析柱纯化后，F（ab′）2片断的回收率为理论值的48％。用双抗体夹心竞争抑制ELISA法测定经S－200纯化的AFPMcAbF（ab′）2片断的活性。其活性与完整的AFPMcAb相同。抑制效价均为1：6400。The preparation of F(ab')2 fragments by digesting Anti-AFP McAb with pepsin need a long time and showed low productivity. However, after being digested by cysteine activated papain in the absence of cysteine for 4 hours, SDS-PAGE showed that over 90% of the McAb was degraded into F(ab')2 fragments. The fragments were purified by Sephacryl-200 column, and the result showed that the recovery rate of thenl was 48% of theoretical value. Double antibody sandwich competitivc inhibition ELISA proved that the activity of ptlrified F(ab')2 fragments was identical with that complete anti-AFP McAb, and both the inhibition titers of them were 1: 6400.

关 键 词：甲胎蛋白 单克隆抗体 F(ab')2片段 木瓜酶 

分 类 号：R392－33[医药卫生—免疫学]