CAJ | 학술논문

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Superdex 200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。
목적：연구중조인첩상신경영양인자（rhCNTF）돌변체적취을이순（PEG）화수식，대rhCNTF적PEG화산물진행초보분리순화급상관생물활성검측。방법：채용분자생물학기술경점돌변득도rhCNTF적돌변체CN10，통과실험설계연구CN10적최가PEG화조건；채용분자사층석방식대우련산물진행초보순화，최후용ELISA화소서체중증장억제법검측PEG화후적CN10단백적생물활성。결과：능운용mPEG-MAL대CN10진행정점수식，PEG화후용Superdex 200능구분리CN10；PEG화후적CN10매2 d복강주사1차，대소서체중적증장억제솔가체50%，여rhCNTF매천주사2차적체중증장억제작용상당。결론：CN10단백재PEG화수식후，기감중효응지속시간명현연장。
Objective: To study the PEGylation of recombinant human ciliary neurotrophic factor(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.