纯化的抗乙酰胆碱酯酶单克隆抗体对乙酰胆碱酯酶活性的影响(英文)  

Effects of purified anti-acetylcholinesterase monoclonal antibodies on the enzyme activity

作　　者：王家蓓[1] 李凤珍[1] 傅风华[1] 孙曼霁[1] 阎国珍 辛颜彬[2] 徐品芳 

机构地区：[1]军事医学科学院药物毒物研究所,北京100850 [2]军事医学科学院微生物及流行病研究所,北京100850

出　　处：《中国药理学与毒理学杂志》1989年第3期225-229,共5页Chinese Journal of Pharmacology and Toxicology

基　　金：中国科学基金资助课题,生85-90~~

摘　　要：本文研究了Balb/c小鼠杂交瘤细胞分泌的抗乙酰胆碱酯酶单克隆抗体对电鳐电器官乙酰胆碱酯酶催化反应的影响及其机理。小鼠腹水抗体经蛋白A琼脂糖亲和层析分离纯化,达到聚丙烯酰胺凝胶电泳纯。酶联免疫吸附试验检测了11株单克隆抗体,滴度在1:5000～1:300 000范围内。在以乙酰胆碱为底物的催化反应中,3F3、2G8及1H11三株杂交瘤分泌的单克隆抗体对乙酰胆碱酯酶活性有明显的抑制作用,抑制作用随单克隆抗体用量的加大而增强。但在以乙酰靛酚为底物的催化反应中,仅3F3显示抑制作用。因而推论,3F3作用在乙酰胆碱酯酶的活力中心,而2G8及1H11可能作用在活力中心的邻近部位.乙酰胆碱酯酶的活力中心不仅是酶的催化部位,而且也是酶分子上的一个抗原决定簇.乙酰胆碱酯酶活力中心具有催化及诱导抗体产生的双重功能.Mouse ascitcs monoclonal anti-bodies were purified by Protein A-Scpharosc affinity chromatography and examined by clcctrophorcsis and ELISA.Studies were undertaken to detect the effects of purified anti-acetylcholinesterase monoclonal antibodies(anti-AChE McAbs)on the catalytic activity of acetylcholincstcrasc from the electric organ of Torpediniforms Nacline timilei.Three out of eleven anti-AChE McAbs,namely,3F3,2G8,1H11,significantly decreased the enzyme activity when acetylcholine was used as the substrate.Further study with indophenol acetate as the substrate showed that among them,only 3F3 caused inhibition to the enzyme activity.It suggests that 3F3 might be bound up with the active center of AChE,while 2G8 and 1H11 presumably be bound up with the sites near the active center.Thus we suppose that the active center of AChE acts as not only the catalytic site but also one of the antigenic determinants of the enzyme.The active center of AChE is bifunctional in nature,catalytic and antibody-inducing.

关 键 词：乙酰胆碱酯酶 单克隆抗体 乙酰胆碱 乙酰靛酚 

分 类 号：Q785[生物学—分子生物学]