Electrochemical DNA sensing using osmium complexes as hybridization indicators

被引:35
作者
Del Pozo, MV
Alonso, C
Pariente, F
Lorenzo, E [1 ]
机构
[1] Univ Autonoma Madrid, Fac Ciencias, Dept Quim Analit & Anal Instrumental, E-28049 Madrid, Spain
[2] Univ Autonoma Madrid, Fac Ciencias, Dept Quim Fis Aplicada, E-28049 Madrid, Spain
关键词
DNA biosensor; hybridization indicators; differential pulse voltammetry; osmium complexes;
D O I
10.1016/j.bios.2004.08.002
中图分类号
Q6 [生物物理学];
学科分类号
071011 ;
摘要
A surface-based method for the study of the interactions of DNA with redox-active osmium complexes is described. The study was carried out using gold electrodes modified with DNA by adsorption and [Os(bpy)(3)](3+/2+) (bpy = 2,2'-bipyridyl) or [Os(phen)(3)](3+/2+) (phen = 1,10-phenantroline) as electrochemical indicators. The method, which is simple and reagent saving, allows the accumulation of osmium complexes on the DNA layer. The amount of osmium complex bound by the layer of double-stranded (dsDNA) or single-stranded DNA (ssDNA) adsorbed at gold electrodes was estimated from the cyclic voltammetric (CV) peak charge of osmium complex reduction. The dissociation constants (K) for the oxidized and reduced forms of a bound species are also estimated. [Os(phen)(3)](3+/2+) was applied to a probe for electrochemical DNA sensing. A thiol-linked single-stranded DNA probe was immobilized through the S-Au bonding to 70 pmol/cm(2) on a gold electrode. Following hybridization with the complementary DNA, the osmium complex was electrochemically accumulated on the double-stranded DNA layer and the differential pulse voltammogram. for this electrode gave an electrochemical signal due to the redox reaction of [Os(phen)3]3+/2+ that was bound to the double-stranded DNA on the electrode. (C) 2004 Elsevier B.V. All rights reserved.
引用
收藏
页码:1549 / 1558
页数:10
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