甲基营养型酵母系统表达的重组人表皮生长因子的纯化及其性质  

Purification of Recombinant hEGF Expressed in Yeast Pichia pastoris and the Study on Its Characters

作　　者：黄秉仁[1] 蔡良婉[1] 王欣[1] 马雪梅[1] 马晓骊[1] 李旌军[1] 李红卓[1] 韦忠明[2] 梁青[2] 郑华军[2] 唐琪浩[2] 唐申秀[2] 廖洪涛[2] 向绪传[2] 

机构地区：[1]中国医学科学院中国协和医科大学基础医学研究所医学分子生物学国家重点实验室,北京100005 [2]Middle-Scale Experimental Base of the Institute of Basic Medical Sciences,CAMS& PUMC and Jin Ke Company

出　　处：《中国医学科学院学报》2001年第2期106-110,共5页Acta Academiae Medicinae Sinicae

基　　金：国家"九五"科技攻关项目! (96-C02-01-04A)&&

摘　　要：目的 获得可用于动物试验和临床试验的人表皮生长因子原料药。方法 化学合成的人表皮生长因子基因在甲基营养型酵母中表达并分泌到培养基中的人表皮生长因子蛋白经 Phenysepharose 6 Fast Flow（ high sub）柱、 Q-sepharose High Performance柱、 Superdex 30柱纯化后进行生物学性质检测。结果 纯化后的蛋白产物纯度达 98％，是无热源、无内毒素、无甲基营养型酵母染色体 DNA污染的，有正确的分子量、等电点、氨基末端氨基酸顺序、肽图、紫外光谱、及生物活性性质的重组蛋白。结论 纯化蛋白符合动物试验和临床试验的要求，为制备多种人表皮生长因子制剂进行动物实验和临床试验打下了基础。Objective To obtain recombinant human epidermal growth factor(hEGF) that can be used in animal experiments and clinical trial. Method Chemically synthesized hEGF gene was expressed in Yeast Pichia pastoris and the secretory hEGF was purified by Phenysepharose 6 Fast Flow（ high sub） ,Q-sepharose High Performance ,and Superdex 30 chromatography， and its characters were studied by respective methods . Results The purified hEGF doesn't contain pyrogen, endotoxin, or yeast chromosome DNA and the purity reached 98％ .The recombinant human EGF has correct molecular weight , pI, N-terminal amino acids sequences ,peptide map ,ultraviolet spectrum and well-biological activity. Conclusion The purified hEGF is in accord with the requirements for animal experiments and clinical trial which provids the basis of preparing EGF agents for clinical test.

关 键 词：重组人表皮生长因子 纯化 化学性质研究 生物活性 甲基营养型酵母系统 

分 类 号：R392－33[医药卫生—免疫学] Q78[医药卫生—基础医学]