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Ilya N. Kurochkin, Alexandra D. Vasilyeva, Evgeniy G. Evtushenko, Arkady V. Eremenko, Dmitry V. Pergushov, Larisa V. Sigolaeva
Enzymes
in the development of physico-chemical methods for biomedical research
Abstract
Abstract.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.
Key words: enzyme biosensors, substrate recirculation, surface
modification, polymers, SERS ELISA, tandem mass spectrometry, enzymatic
hydrolysis
Copyright (C) Chemistry Dept., Moscow State University, 2002
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