中华医学超声杂志

应用化学 ›› 2009, Vol. 26 ›› Issue (6): 637-641.

• 研究论文 • 上一篇    下一篇

采用SFAD将碳酸酐酶偶联至无活性官能团聚合物表面的新方法

王琴梅1,张涤华2,张静夏3   

  1. 1. 广州市中山大学附属一院生物活性材料研究室
    2. 中山大学附属第一医院血透中心
    3. 中山大学药学院
  • 收稿日期:2008-06-20 修回日期:2008-09-16 出版日期:2009-06-10 发布日期:2009-06-10
  • 通讯作者: 王琴梅

A New Method of Immobilization of Carbonic Anhydrase onto Polymer Surface without Active Functional Groups by Using SFAD as a Crosslinker

  • Received:2008-06-20 Revised:2008-09-16 Published:2009-06-10 Online:2009-06-10

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摘要：

摘 要 采用异双官能团光偶联剂－Sulfosuccinimidyl (perfluoroazidobenzamido) ethyl-1,3-dithio propionate，SFAD)，将碳酸酐酶（Carbonic anhydrase，CA)接枝在无活性官能团聚合物－聚甲基戊烯（Polymethyl-pentene，PMP)膜式氧合器表面，以对硝基苯酚乙酸酯(p-nitrophenyl acetate,p-NPA)为底物，采用紫外分光光度计测定了接枝CA的活性、浓度、重复利用性、储存稳定性。结果表明，采用SFAD为交联剂能将酶成功地偶联在无活性官能团聚合物表面，在保持酶活性的同时获得较高的接枝效率；共价接枝CA（Covalently immobilized CA，CACI）的浓度随SFAD浓度的增加而增加，最大可达理论单分子层接枝量的85.86％；CACI比物理吸附的CA（Physically adsorbed CA，CAPA）具更好的重复利用性；37°C下, CACI 比CA溶液表现出更好的储存稳定性。

关键词: 聚甲基戊烯, 光偶联剂, 碳酸酐酶, 对硝基苯酚乙酸酯

Abstract:

ABSTRACT Carbonic anhydrase(CA) was immobilized onto the surface of hollow fiber membrane of polymethyl pentene(PMP) without any active functional groups by using sulfosuccinimidyl [perfluoroazidobenzamido] ethyl-1,3-dithiopropionate (SFAD), a photoactivatable heterobifunctional crosslinker. Using p-nitrophenyl acetate (p-NPA) as a substrate, the activity, concentration, storage stability and re-usability of immobilized CA on PMP hollow fibers were studied by ultraviolet spectrophotometer. The preliminary data showed that CA could be successfully introduced onto PMP surface with catalysis activity and immobilization efficiency.The activity of covalently immobilized CA increased with the increase of concentration of SFAD, and the maximum was 85.86% of the theoretical activity of CA spread on PMP surface in monolayer in studied range. Covalently immobilized CA showed higher reusability compared to physically adsorbed CA, and higher storage stability compared to free CA in solution at 37℃.

Key words: Polymethyl pentene, Photoreactive crosslinker, Carbonic anhydrase, p-nitrophenyl acetate

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