酶在生物医学研究中发展的理化方法

Ilya N. Kurochkin, Alexandra D. Vasilyeva, Evgeniy G. Evtushenko, Arkady V. Eremenko, Dmitry V. Pergushov, Larisa V. Sigolaeva
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引用次数: 0

摘要

本文讨论了伏安法,表面增强拉曼光谱(SERS)和质谱法与酶催化串联的可能性。介绍了氧化酶和脱氢酶电化学生物传感器的基本工作原理。研究表明,利用循环酶反应和底物循环的生物传感器具有最佳的灵敏度。由于使用聚合物对电极表面进行有效修饰和酶的非破坏性固定化,生物传感器分析的分析潜力得到了显著改善。数据显示如何使用酶标签扩大生物分析SERS的应用范围。利用SERS高灵敏度测量酶标记(过氧化物酶、碱性磷酸酶、β-半乳糖苷酶)活性的可能性,实际上为开发测定各种抗原的方法开辟了一个新的通用平台。以最常用的蛋白酶为例,讨论了质谱法蛋白质组学研究方法发展的主要趋势,以及蛋白酶在质谱实验设计中的作用。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
ENZYMES IN THE DEVELOPMENT OF PHYSICO-CHEMICAL METHODS FOR BIOMEDICAL RESEARCH
The paper addresses the possibilities of voltammetry, surface-enhanced Raman spectroscopy (SERS) and mass spectrometry in tandem with enzymatic catalysis. The basic principles of operation of electrochemical biosensors based on oxidases and dehydrogenases are described. It has been shown that biosensors using cyclic enzymatic reactions and substrate recycling have the best sensitivity. The variants of significant improvement of the analytical potential of biosensor analysis due to the use of polymers for effective modification of the electrode surface and non-destructive immobilization of enzymes are illustrated. The data demonstrating how the use of enzyme labels expands the range of bioanalytical applications of SERS are presented. The possibility of highly sensitive measurement of the activity of enzyme labels (peroxidase, alkaline phosphatase, β-galactosidase) using SERS, in fact, opens up a new universal platform for the development of methods for the determination of various antigens. By way of example of the most commonly used proteases, the main trends in the development of the methodology of proteomic studies by mass spectrometry, as well as the role of proteases in the design of mass spectrometric experiments, are considered.
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