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›› 2006, Vol. 6 ›› Issue (4): 619-622.

• 生化工程专栏 • 上一篇    下一篇

不同分子量聚乙二醇单修饰重组人干扰素a-2a

张霖琳,郑春杨,雷建都,马光辉,苏志国   

  1. 中国科学院过程工程所生化工程国家重点实验室
  • 出版日期:2006-08-20 发布日期:2006-08-20

Preparation and Characterization of the rhIFN-a-2a Modified by Polyethylene Glycol with Different Molecular Weights

ZHANG Lin-lin,ZHENG Chun-yang,LEI Jian-du,MA Guang-hui,SU Zhi-guo   

  1. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering
  • Online:2006-08-20 Published:2006-08-20

摘要: 利用N-羟基琥珀酰亚胺活化制备不同分子量的聚乙二醇修饰剂(NHS-mPEG,分子量5000, 10000, 20000),考察三者水解动力学性质的差异,结合正交实验确定了不同分子量修饰剂制备单条PEG链修饰重组人干扰素a-2a(rhIFN-a-2a)的最佳反应条件,用离子交换法对产物进行分离纯化. 研究了不同分子量聚乙二醇干扰素结合物的体外活性,比较其蛋白收率. 实验结果表明,修饰剂分子量增大,应选择蛋白与修饰剂的反应活性高的修饰反应条件,体外活性虽然降低,但同时单修饰聚乙二醇干扰素结合物收率更高,产品质量更易控制.

关键词: 重组人干扰素a-2a, 聚乙二醇, 化学修饰

Abstract: Monomethoxy polyethylene glycol with different molecular weights (mPEG5000, 10000 and 20000) were prepared using activated N-hydroxysuccinimede (NHS). NHS-mPEG was investigated by hydrolysis kinetics. Through the experiments based on orthogonal design, the optimized reaction conditions could be proposed to obtain a single PEG chain attached onto the rhIFN-a-2a. Ion exchange chromatography was used to separate and purify PEGylated rhIFN-α-2a products from unPEGylated rhIFN-α-2a. PEGylated rhIFN-a-2a with different molecular weights was contrasted by in vitro bioactivity and protein recovery rate. It was revealed that with the increase of molecular weight of NHS-mPEG, the modification reaction conditions were higher and the in vitro bioactivity was lower, but the protein recovery rate was increased.

Key words: rhIFN-a-2a, polyethylene glycol, chemical modification