中国药理学与毒理学杂志2002，Vol.16Issue(6)：454-458,5.
基因工程人丁酰胆碱酯酶的纯化及其在体外对琥珀酰胆碱的解毒作用
Purification of genetically engineered human butyrylcholinesterase and detoxification of succinylcholine in vitro
摘要
Abstract
AIM To develop a plenty and secure resource of human butyrylcholinesterase, and examine its detoxification efficacy. METHODS The purification of recombinant human butyrylcholinesterase(rhBChE) was based on the fractionation of the hemolymph proteins of silkworm larvae by ammonium sulfate and the procainamide-Sepharose 4B affinity chromatography in batches. RESULTS The overall yield of the SDS-PAGE pure rhBChE activity came to 66% of the hemolymph. The specific enzyme activity of the purified rhBChE reached at 715 mmol*min-1*g-1 protein. The catalytic activity of the enzyme stored in 20% glycerol, 0.02% NaN3, 10 mmol*L-1 sodium phosphate buffer (pH 7.4) at -20℃ for one year showed little change. Mice challenged ip with 1.5 LD of succinylcholine preincubated with rhBChE survived (18/18) without any symptom and sign of intoxication, whereas the mice in the succinylcholine control group all (18/18) died of convulsion and suffocation within 2-5 min. CONCLUSION The methods used are valid for purification of rhBChE from the hemolymph of the infected larvae. The genetically engineered rhBChE is of potential use in the therapy of succinylcholine crisis.关键词
丁酰胆碱酯酶/蚕,家蚕/琥珀酰胆碱/色谱法,亲和Key words
butyrylcholinesterase/silkworms, Bombyx mori/succinylcholine/chromatography, affinity分类
医药卫生引用本文复制引用
魏婉丽,徐海,池木根,李凤珍,秦浚川,孙曼霁..基因工程人丁酰胆碱酯酶的纯化及其在体外对琥珀酰胆碱的解毒作用[J].中国药理学与毒理学杂志,2002,16(6):454-458,5.基金项目
"九五"全军医药基金资助项目(96-Z-016) （96-Z-016）
