Hayat Akhtar, Sassolas Audrey, Rhouati Amina, Marty Jean-Louis
IMAGES EA 4218, University of Perpignan, Perpignan Cedex, France.
Methods Mol Biol. 2013;1051:209-16. doi: 10.1007/978-1-62703-550-7_13.
This paper describes a novel, simple, and versatile protocol for covalent immobilization of enzyme on electrode. The immobilization method is based on the combination of diazonium salt electrografting and click chemistry. The ethynyl-terminated monolayers are obtained by diazonium salt electrografting, then, in the presence of copper (I) catalyst, the ethynyl modified surfaces reacts efficiently and rapidly with enzyme bearing an azide function (azido-enzyme), thus forming a covalent 1,2,3-triazole linkage by means of click chemistry. The ethynyl-terminated film preserves the activity of the immobilized enzyme. The click chemistry along with binary film of diazonium salts offers a variety of good characteristics including high sensitivity, good repeatability and reusability, rapid response and long term stability of the system. Thus, because of the chemoselective reactivity and quantitative yield of the click reaction, an ethynyl-terminated monolayer can be treated as a general platform for obtaining reliable coverage of a wide range of azido-terminated species of interest for various sensing applications.
本文描述了一种用于将酶共价固定在电极上的新颖、简单且通用的方案。该固定方法基于重氮盐电接枝和点击化学的结合。通过重氮盐电接枝获得乙炔基封端的单分子层，然后，在铜（I）催化剂存在下，乙炔基修饰的表面与带有叠氮化物功能的酶（叠氮基酶）高效快速地反应，从而通过点击化学形成共价的1,2,3-三唑键。乙炔基封端的膜保留了固定化酶的活性。点击化学与重氮盐二元膜一起提供了多种良好特性，包括高灵敏度、良好的重复性和可重复使用性、快速响应以及系统的长期稳定性。因此，由于点击反应的化学选择性反应性和定量产率，乙炔基封端的单分子层可被视为一个通用平台，用于获得各种传感应用中广泛感兴趣的叠氮基封端物种的可靠覆盖。
