重组人睫状神经营养因子的聚乙二醇化修饰  

PEGylation of Recombinant Human Ciliary Neurotrophic Factor

作　　者：梁凌宇[1] 雷清[1] 高雪峰[1] 杨军[1] 应莲芳[1] 蒋琳[1] 

机构地区：[1]兰州生物制品研究所有限责任公司,甘肃兰州730046

出　　处：《生物技术通讯》2014年第1期65-67,共3页Letters in Biotechnology

摘　　要：目的:研究重组人睫状神经营养因子(rhCNTF)突变体的聚乙二醇(PEG)化修饰,对rhCNTF的PEG化产物进行初步分离纯化及相关生物活性检测。方法:采用分子生物学技术经点突变得到rhCNTF的突变体CN10,通过实验设计研究CN10的最佳PEG化条件;采用分子筛层析方式对偶联产物进行初步纯化,最后用ELISA和小鼠体重增长抑制法检测PEG化后的CN10蛋白的生物活性。结果:能运用mPEG-MAL对CN10进行定点修饰,PEG化后用Superdex200能够分离CN10;PEG化后的CN10每2 d腹腔注射1次,对小鼠体重的增长抑制率可达50%,与rhCNTF每天注射2次的体重增长抑制作用相当。结论:CN10蛋白在PEG化修饰后,其减重效应持续时间明显延长。Objective： To study the PEGylation of recombinant human ciliary neurotrophic faetor（rhCNTF）, estab lish the preliminary separation and purification of the polyethylene glycol-modified rhCNTF, and then to detect the biological activity of PEGylation rhCNTF. Methods： Using molecular biology techniques to obtain the mutants CN10 of rhCNTF by point mutation, then to study the conditions of PEG conjugation by design of experiment. The prod uct of PEGylation rhCNTF were purified by molecular sieve chromatography, finally the biological activity were de tected by ELISA and the method of weight loss in normal mouse. Results： CN10 can be successfully fixed-point modified by mPEG-MAL, the weight loss in mice experiments showed that the weight increase inhibitory rate of PEGylation CN10 were up to 50% when received an intraperitoneal injection every other day, it was equivalent to inhibitory effect of rhCNTF which injected twice a day. Conclusion： After PEG modification, the biological effect of rhCNTF was significantly prolonged.

关 键 词：人睫状神经营养因子 聚乙二醇化修饰 纯化 体重增长抑制 

分 类 号：Q78[生物学—分子生物学] Q502