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학술지 Multienzyme-Modified Biosensing Surface for the Electrochemical Analysis of Aspartate Transaminase and Alanine Transaminase in Human Plasma
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저자
한용덕, 송승연, 이준황, 이대식, 윤현철
발행일
201112
출처
Analytical and Bioanalytical Chemistry, v.400 no.3, pp.797-805
ISSN
1618-2642
출판사
Springer
DOI
https://dx.doi.org/10.1007/s00216-011-4797-6
초록
We investigated the electrochemical detection of aspartate transaminase (AST) and alanine transaminase (ALT) by using a multienzyme-modified electrode surface. Determination of the activities of transaminases in human serum is clinically significant because their concentrations and ratios indicate the presence of hepatic diseases or myocardial dysfunction. For electrochemical detection of AST and ALT, enzymes that participate in the reaction mechanism of AST and ALT, such as pyruvate oxidase (POX) and oxaloacetate decarboxylase, were immobilized on an electrode surface by using an amine-reactive self-assembled monolayer and a homobifunctional cross-linker. In the presence of suitable substrates such as l-aspartate (l-alanine) and 慣-ketoglutarate, AST and ALT generate pyruvate as an enzymatic end product. To determine the activities of AST and ALT, electroanalyses of pyruvate were conducted using a POX and ferrocenemethanol electron shuttle. Anodically generated oxidative currents from multienzyme-mediated reactions were correlated to AST and ALT levels in human plasma. On the basis of the electrochemical analysis, we obtained calibration results for AST and ALT concentrations from 7.5 to 720 units/L in human plasma-based samples, covering the required clinical detection range. [Figure not available: see fulltext.] © 2011 Springer-Verlag.
키워드
Clinical enzyme analysis, Electrochemical biosensor, Immobilized enzyme surface, Multienzyme assay
KSP 제안 키워드
Clinical detection, Electrochemical biosensor, Electrochemical detection(ECD), Electrode surface, Enzyme surface, Human serum, Immobilized enzyme, L-Aspartate, Oxaloacetate decarboxylase, Pyruvate oxidase(POD), Reaction mechanisms