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抗丁氏双鳍电鳐电器官乙酰胆碱酯酶(AChE)的抑制性单克隆抗体IH11(IgM),丁氏双鳍电鳐,石纹电线电器官,比目鱼肌肉,大鼠脑,牛脑,人脑,牛红细胞及人红细胞ACbE均以不同的亲和层析法提纯。用酶抗原免疫测定法及酶联免疫吸附测定法检测了1H11与上述脊椎动物的AChE的交叉免疫反应性.结果表明,1H11可与石纹电绥,比目鱼肌肉,大鼠脑、牛脑及人脑AChE有交叉免疫反应。显示了1H11所针对的抗原决定簇在进化中的高度保守性。1H11与人,牛红细胞AChE及人丁酰胆碱酯酶(BChE)无交叉免疫反应性。提示哺乳类脑AChE与红细胞AChE及人BChE有不同的抗原决定簇。1H11对电鳐及电鳗AChE活性产生明显抑制。但对其它几个脊椎动物的AChE活性无抑制.表明1H11所针对的抗原决定簇在电鳐AChE的三级结构中靠近其活性中心,而在其它脊椎动物的AChE中刚远离其活性中心.AChE阴离子部位抑制剂依酚氟铵不影响抗原抗体反应,表明1H11不结合在阴离子部位。N-糖苷酶F处理使ACbE去糖基化后。1H11不再能与AChE结合,表明至少1H11所识别的抗原决定簇的主要部分是N-多醣.抑制性单克隆抗体1H11对脑ACh?
Anti-Dichotoma bifidus Electrical Potential Inhibitors of AChE Monoclonal Antibodies IH11 (IgM), Dichotomous Dichotomous, Staurotline Electrical, Soleus Muscle, Rat Brain, Bovine Brain, Human Brain, bovine red blood cells and human erythrocyte ACbE were purified by different affinity chromatography. The cross-reactivity of 1H11 with AChE of vertebrates was tested by enzyme immunoassay and enzyme-linked immunosorbent assay (ELISA). The results showed that 1H11 could cross-react with AChE, Ishikari, soleus muscle, rat brain, bovine brain and human brain. The antigenic determinants against 1H11 are highly conserved in evolution. 1H11 has no cross-reactivity with human, bovine red blood cell AChE and human butyrylcholinesterase (BChE). Tip mammalian brain AChE and red blood cells AChE and human BChE have different antigenic determinants. 1H11 significantly inhibited the activity of AChE in electric and electric eels. But did not inhibit AChE activity of several other vertebrates. Indicating that the epitope targeted by 1H11 is close to its active site in the tertiary structure of AChE, and far away from its active site in AChE of other vertebrates. AChE anion site inhibitor phenphenamine did not affect the antigen-antibody reaction, indicating that 1H11 does not bind to the anion site. After N-glycosidase F treatment deglycosylated ACbE. 1H11 no longer binds to AChE, indicating that at least the major portion of the epitope recognized by 1H11 is N-polysaccharide. Inhibitory monoclonal antibody 1H11 on brain ACh?