色谱 ›› 2015, Vol. 33 ›› Issue (11): 1121-1125.DOI: 10.3724/SP.J.1123.2015.06028

• 特殊选择性分离介质的制备专栏 • 上一篇    下一篇

蛋白质组学中的固定化酶反应器制备的研究进展

张凌怡1, 王兵兵1, 上官露露1, 张润生2, 陈建虎1, 张维冰1   

  1. 1. 上海市功能性材料化学重点实验室, 华东理工大学, 上海 200237;
    2. 上海市现场物证重点实验室, 上海市刑事科学技术研究院, 上海市公安局, 上海 200083
  • 收稿日期:2015-06-17 出版日期:2015-11-08 发布日期:2012-05-30
  • 通讯作者: 张维冰
  • 基金资助:

    国家重大科学仪器设备开发专项(2012YQ120044);上海市刑事科学技术研究院开放课题.

Development of preparation of immobilized enzyme reactors in proteomics

ZHANG Lingyi1, WANG Bingbing1, SHANGGUAN Lulu1, ZHANG Runsheng2, CHEN Jianhu1, ZHANG Weibing1   

  1. 1. Shanghai Key Laboratory of Functional Materials Chemistry, East China University of Science and Technology, Shanghai 200237, China;
    2. Shanghai Key Laboratory of Crime Scene Evidence, Shanghai Research Institute of Criminal Science and Technology, Shanghai Public Security Bureau, Shanghai 200083, China
  • Received:2015-06-17 Online:2015-11-08 Published:2012-05-30

摘要:

固定化酶反应器作为蛋白质组学研究中"bottom-up"策略重要的组件,具有酶解快速、酶解效率高、酶稳定性和活性高、简单易操作、能够与多种检测方式联用等优点,对于发展高效快速的蛋白质组学分析方法具有重要意义。本文就固定化酶反应器的制备方法及其在蛋白质组学中的应用做简单的概述,着重介绍酶的固定方法、固定化酶的载体、用于固定的酶的种类。近几年固定化酶反应器的研究集中于提高固酶量、保持酶活性、增加酶解效率、减小非特异性吸附等方面。研究结果表明,采用纳米材料、整体材料等新型载体,提高载体亲水性,采用多酶同时酶解等方法能够有效改善固定化酶反应器的性能,提高蛋白质的鉴定效率。

关键词: 蛋白质组学, 固定化酶反应器, 酶, 载体材料, 制备

Abstract:

As an important part in "bottom-up" strategy of proteomics, immobilized enzyme reactors have great significance in the development of fast and more efficient protein analytical method, owing to its advantages of high speed and enzymatic efficiency, good stability and activity, easy operation, and the possibility of hyphenating with multiple detection instruments. In this paper, the preparation methods of immobilized enzyme reactors and their applications in proteomic investigation are introduced, focusing on the nature of enzymes, the immobilization methods and the carrier materials used for immobilizing enzyme. In recent years, the investigations are focused on increasing the immobilization amounts of enzyme, keeping enzymatic activity, improving enzymatic efficiency and decreasing nonspecific adsorption. The investigation results showed that by using novel carriers such as nanomaterial and monolith, increasing of hydrophilicity of carrier and tandem hydrolysis with multiple enzymes can greatly improve the performance of immobilized enzyme reactors and increase protein identification efficiencies.

Key words: carrier material, enzyme, immobilized enzyme reactors, preparation, proteomics

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