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›› 2006, Vol. 6 ›› Issue (6): 954-958.

• 生化工程专栏 • 上一篇    下一篇

多孔聚(甲基丙烯酸环氧丙酯-二乙烯基苯)微球的改性及作为蛋白质疏水层析介质的应用

张颖,王仁伟,马光辉,谭天伟,苏志国   

  1. 中国科学院过程工程研究所生化工程国家重点实验室
  • 出版日期:2006-12-20 发布日期:2006-12-20

Modification of Poly(Glycidyl Methacrylate-Divinylbenzene) Porous Microspheres with Polyethylene Glycol and Their Use as HIC Medium for Protein Adsorption

ZHANG Ying,WANG Ren-wei,MA Guang-hui,TAN Tian-wei,SU Zhi-guo   

  1. State Key Laboratory of Biochemical Engineering, Institute of Process Engineering, Chinese Academy of Sciences
  • Online:2006-12-20 Published:2006-12-20

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摘要： 用悬浮聚合方法合成了富含环氧基团的多孔聚(甲基丙烯酸环氧丙酯-二乙烯基苯) [Poly(Glycidyl Methacrylate-Divinylbenzene), P(GMA-DVB)]微球，为了消除其对蛋白质的不可逆吸附，探索用聚乙二醇(Polyethylene Glycol, PEG)对微球进行改性，制备了带有PEG配基的P(GMA-DVB)微球. 实验考察了反应条件对PEG固载量的影响规律，发现最适反应温度为80℃. 以牛血清白蛋白(Bovine Serum Albumin, BSA)和胰蛋白酶作为模型蛋白，考察了PEG改性前后P(GMA-DVB)微球对蛋白质的吸附性能. 改性前P(GMA-DVB)微球有30%~40%的不可逆吸附，蛋白质回收率仅为60%~70%. 改性后介质消除了不可逆吸附，对蛋白质的吸附作用表现为可逆的疏水相互作用，吸附BSA和胰蛋白酶的质量回收率和活性回收率都在97%以上. 结果表明，PEG-P(GMA-DVB)微球可以作为一种新型介质进一步应用于蛋白质的吸附与分离.

关键词: 聚甲基丙烯酸环氧丙酯微球, 改性, 聚乙二醇, 蛋白质吸附, 疏水相互作用

Abstract: Porous poly(glycidyl mathacrylate-divinylbenzene) [P(GMA-DVB)] microspheres containing epoxy groups were synthesized through suspension polymerization. In order to minimize non-reversible adsorption of proteins, polyethylene glycol (PEG) was coupled onto the microspheres via the reaction with epoxy groups. The reaction process was studied and optimized with respect to the amount of PEG coupling. The optimum temperature was 80℃. Bovine serum albumin (BSA) and trypsin were used as two model proteins to examine adsorption and desorption properties of the modified P(GMA-DVB) microspheres. The modified microspheres had little non-reversible adsorption for BSA and trypsin. The mass recovery rate and activity recovery rate were more than 97%. The adsorption of protein on the modified microspheres was reversible hydrophobic interaction. The modified microspheres can be used for hydrophobic interaction chromatography of proteins.

Key words: poly glycidyl mathacrylate microspheres, modification, glycol ethylene, protein adsorption, hydrophobic interaction medium