中国医学科学院学报
中國醫學科學院學報
중국의학과학원학보
ACTA ACADEMIAE MEDICINAE SINICAE
2001年
2期
106-110
,共5页
黄秉仁%蔡良婉%王欣%马雪梅%马晓骊%李旌军%李红卓%韦忠明%梁青%郑华军%唐琪浩%唐申秀%廖洪涛%向绪传
黃秉仁%蔡良婉%王訢%馬雪梅%馬曉驪%李旌軍%李紅卓%韋忠明%樑青%鄭華軍%唐琪浩%唐申秀%廖洪濤%嚮緒傳
황병인%채량완%왕흔%마설매%마효려%리정군%리홍탁%위충명%량청%정화군%당기호%당신수%료홍도%향서전
重组人表皮生长因子纯化化学性质研究生物活性
重組人錶皮生長因子純化化學性質研究生物活性
중조인표피생장인자순화화학성질연구생물활성
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。
目的穫得可用于動物試驗和臨床試驗的人錶皮生長因子原料藥。方法化學閤成的人錶皮生長因子基
因在甲基營養型酵母中錶達併分泌到培養基中的人錶皮生長因子蛋白經Phenysepharose 6 Fast Flow(high sub)
柱、Q-sepharose High Performance 柱、Superdex 30 柱純化後進行生物學性質檢測。結果純化後的蛋白產物純度達98％，是無熱源、無內毒素、無甲基營養型酵母染色體DNA汙染的，有正確的分子量、等電點、氨基末耑氨基痠順序、肽圖、紫外光譜、及生物活性性質的重組蛋白。結論純化蛋白符閤動物試驗和臨床試驗的要求，為製備多種人錶皮生長因子製劑進行動物實驗和臨床試驗打下瞭基礎。
목적획득가용우동물시험화림상시험적인표피생장인자원료약。방법화학합성적인표피생장인자기
인재갑기영양형효모중표체병분비도배양기중적인표피생장인자단백경Phenysepharose 6 Fast Flow(high sub)
주、Q-sepharose High Performance 주、Superdex 30 주순화후진행생물학성질검측。결과순화후적단백산물순도체98％，시무열원、무내독소、무갑기영양형효모염색체DNA오염적，유정학적분자량、등전점、안기말단안기산순서、태도、자외광보、급생물활성성질적중조단백。결론순화단백부합동물시험화림상시험적요구，위제비다충인표피생장인자제제진행동물실험화림상시험타하료기출。
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 se
quences, 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 prepar ing EGF agents for clinical test.
