Electrochemical detection for horseradish peroxidase-based enzyme immunoassay using p-aminophenol as substrate and its application in detection of plant virus

Citation
W. Sun et al., Electrochemical detection for horseradish peroxidase-based enzyme immunoassay using p-aminophenol as substrate and its application in detection of plant virus, ANALYT CHIM, 434(1), 2001, pp. 43-50
Citations number
19
Language
INGLESE
art.tipo
Article
Categorie Soggetti
Spectroscopy /Instrumentation/Analytical Sciences
Journal title
ANALYTICA CHIMICA ACTA
ISSN journal
0003-2670 → ACNP
Volume
434
Issue
1
Year of publication
2001
Pages
43 - 50
Database
ISI
SICI code
0003-2670(20010425)434:1<43:EDFHPE>2.0.ZU;2-D
Abstract
A sensitive electrochemical method for horseradish peroxidase (HRP)-based e nzyme immunoassay using p-aminophenol (PAP) as substrate is described. The enzymatic product of PAP oxidation with H2O2 catalyzed by HRP in Britton-Ro binson (B-R) buffer solution of pH 4.7 is 3,4-di-[(4-hydroxyphenyl)amino]-6 -[(4-hydroxyphenyl) imino]-2,4-cyclohexadiene-1-one, which yields a sensiti ve second order derivative linear-sweep voltammetric peak at potential of - 0.56 V (versus Ag/AgCl) in B-R buffer solution of pH 7. The processes of th e enzymatic catalyzed reaction and the electrode reduction of the enzymatic product have been carefully investigated. By using this voltammetric respo nse, HRP can be measured with a detection Limit of 0.4 mUI(-1) and a linear range of 1-100 mUI(-1). So it was further applied to immunoassay and cucum ber mosaic virus (CMV) was detected as a model. The detection limit of the purified CMV is 0.5 ng ml(-1), which is 10 times lower than that of traditi onal colorimetric o-phenylenediamine (OPD) enzyme-linked immunosorbent assa y (ELISA) method. The results show greatly increased sensitivity by electro chemical enzyme immunoassay. (C) 2001 Elsevier Science B.V. AU rights reser ved.