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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.
In this paper, Balb / c mouse hybridoma cells secreted anti-acetylcholinesterase monoclonal antibody on the electrical organ acetylcholinesterase catalytic reaction and its mechanism. Mouse ascites antibody by Protein A Sepharose affinity chromatography purification, reached polyacrylamide gel electrophoresis pure. Eleven monoclonal antibodies were detected by enzyme-linked immunosorbent assay (ELISA) with a titer ranging from 1: 5000 to 1: 300 000. Monoclonal antibodies secreted by the three hybridomas of 3F3, 2G8 and 1H11 significantly inhibited the activity of acetylcholinesterase (AChE) in the catalytic reaction of acetylcholine substrate. The inhibition increased with the increase of the amount of monoclonal antibody. However, in the catalytic reaction using acetyl-indophenol as a substrate, only 3F3 showed an inhibitory effect. Therefore, it is concluded that 3F3 acts on the center of activity of acetylcholinesterase and that 2G8 and 1H11 may act in the vicinity of the center of activity of acetylcholinesterase, which is not only the catalytic site of the enzyme, but also an antigenic determinant on the enzyme molecule. Acetylcholinesterase activity centers have dual functions of catalyzing and inducing antibody production.