转基因羊奶中促红细胞生成素的提纯  

Purification of enthropoietin from transgenic goat milk

作　　者：汪燕芳[1] 彭进 赵建阳 倪亚明[1] 成国祥 

机构地区：[1]同济大学生命科学与技术学院,上海200092 [2]上海转基因研究中心,上海201203

出　　处：《中国生化药物杂志》2006年第4期215-218,共4页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的从pBLG-hEPO转基因母山羊的羊乳中提纯促红细胞生成素(EPO)。方法经过酸沉淀预处理、SPSepharose FF阳离子交换树脂、butyl FF 16/10疏水交换树脂、Superdex 75 26/60分子筛色谱4步分离,从转基因羊乳中纯化出EPO。结果经SDS-PAGE检测,得到的EPO相对分子质量约为32 000,全程回收率为10.1%,ELISA和蛋白质印迹实验证明其具有天然EPO抗原性。结论此工艺可用于快速分离转基因羊乳中EPO。Purpose The EPO was purified from the milk of pBLG-hEPO transgenic goats,Methods The purification process was composed of four steps ： pre-treatment by acid, SP Sepharose FF ion-exchange, butyl FF 16/10 hydrophobic chromatography and Superdex 75 26/60 molecule sieve chromatography. Results Its molecular weight was 32 000 determined by SDS-PAGE. The total recovery of EPO activity was 10.1%, It showed the same immunity as the natural EPO proved by ELISA and protein western-blotting. Conclusion The process can be used to purify the EPO from the transgenic goat milk.

关 键 词：重组人红细胞生成素 提纯 色谱 

分 类 号：Q513[生物学—生物化学] R284.2[医药卫生—中药学]