睫状神经营养因子突变体的中试工艺研究  

Study on the pilot-scale production of ciliary neurotrophic factor mutant

作　　者：毕华[1] 袁力勇[1] 赵阳[1] 史新昌[1] 李永红[1] 饶春明[1] 王军志[1] 

机构地区：[1]中国药品生物制品检定所生化室,北京100050

出　　处：《中国生化药物杂志》2007年第4期236-239,共4页Chinese Journal of Biochemical Pharmaceutics

摘　　要：目的进行睫状神经营养因子及其突变体的应用开发,研究其中试生产工艺。方法摸索了不同的发酵条件以确定最佳的发酵工艺,采用稀释复性-离子交换色谱-分子筛相结合的方法对其进行复性和纯化。结果得到的蛋白经HPLC检测纯度在95%以上,比活性的达到2.0×106U/mg,末端的15个氨基酸序列和Western blot的结果与要求相符。结论中试工艺简单、迅速,所需设备廉价,有利于大规模生产。Purpose To develop the production of ciliary neurotrophic factor （CNTF） mutant on the pilotscale and to study its pilot process. Methods Exploring many different fermentation conditions to determine optimal process, and using the method of diluted renaturation - ion exchnge chromatography - gel filtration chromatograpgy to renature and purify. Results The purity of final product was over 95 % by HPLC, and the specific activity reached 2.0×10^6U/mg protein. The results of N-terminal 15aa sequence and Western blot were identical with those required. Conclusion The whole pilot procedure was simple and fast, and the equipment needed was cheap, so it was promising to enlarge development.

关 键 词：睫状神经营养因子 突变体 纯化 

分 类 号：TQ464[化学工程—制药化工]