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重组人睫状神经因子聚乙二醇化条件的优化 被引量：1

Condition optimization for modification of recombinant human ciliary neurotrophic factor with polythylene glycol
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摘要 目的对单甲氧基聚乙二醇丁醛(mPEG-ButyrALD,相对分子质量20×103)修饰重组人睫状神经因子(rh-CNTF)的条件进行优化。方法优化了pH值、温度、时间、rh-CNTF与PEG的修饰比例、rh-CNTF浓度等修饰条件,并对修饰产物行分离纯化。鸡胚背根节培养法检测和比较rh-CNTF和PEG-rhCNTF的体外生物学活性。结果rh-CNTF与PEG的修饰比例、rh-CNTF的浓度和pH值是主要影响因素,最佳pH为6.5,最佳rh-CNTF浓度为1～2 mg/ml,rh-CNTF与PEG的最佳质量比为1∶1,时间和温度影响甚微。rh-CNTF的体外神经营养活性约为PEG-rhCNTF的2倍。结论本法优化了修饰重组人睫状神经因子的条件。 Objective To optimize the modification condition of recombinant human ciliary neurotrophic factor （rh-CNTF） with mPEG-ButyrALD （relative molecular weight 20× 10^3 ）. Methods Different pH values, temperatures, times and the mass ratio of rh-CNTF and PEG, the concentration of rh-CNTF was used to obtain better rh-CNTF PEGylation. And then the modified rh-CNTF was separated and purified. The in vitro biological activity of rh-CNTF and PEG rhCNTF was measured by chicken embryo dorsal root ganglion （DRG） culture. Results The mass ratio of rh-CNTF to PEG, rh-CNTF concentration and pH value showed significant effect on rh-CNTF PEGylation, while time and temperature had little influence on the modification effect. The optimal pH value was 6.5, concentration of rh-CNTF was 1 to 2 mg/ml, and mass ratio of rh-CNTF and PEG was 1 ： 1 on modification condition. The in vitro biological activity of rh-CNTF was about 2 times higher than that of PEG-rhCNTF in chicken embryo dorsal root ganglion （DRG） culture. Conclusion Optimized parameters is obtained to modify rh-CNTF with mPEG-ButyrALD.
作者 王彦 范开 夏永鹏 张继兰 李秀梅 邱宗荫
机构地区 重庆医科大学药学院 重庆理工大学化学与生物工程学院制药工程系 重庆市卫生局 重庆市药品检验所生化与生物制品室
出处 《第三军医大学学报》 CAS CSCD 北大核心 2009年第14期1345-1348,共4页 Journal of Third Military Medical University
基金 重庆市自然科学基金(2007BB5254)~~
关键词 聚乙二醇修饰 重组人睫状神经因子 优化 PEGylation recombinant human ciliary neurotrophie factor optimization
分类号 R914 [医药卫生—药物化学] R966 [医药卫生—药理学]
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参考文献10

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二级参考文献9

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第三军医大学学报
2009年 第14期

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