Electrochemiluminescent array to detect oxidative damage in ds-dna using [os(bpy)2(phen-benz-cooh)]2+/nafion/graphene films
Bist, Itti Song, Boya Mosa, Islam M. Keyes, Tia E. Martin, Aaron Forster, Robert J. Rusling, James F.
Bist, Itti
Song, Boya
Mosa, Islam M.
Keyes, Tia E.
Martin, Aaron
Forster, Robert J.
Rusling, James F.
Publication Date
2016-03-25
Type
Article
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Bist, Itti; Song, Boya; Mosa, Islam M. Keyes, Tia E.; Martin, Aaron; Forster, Robert J.; Rusling, James F. (2016). Electrochemiluminescent array to detect oxidative damage in ds-dna using [os(bpy)2(phen-benz-cooh)]2+/nafion/graphene films. ACS Sensors 1 (3), 272-278
Abstract
Reactive oxygen species (ROS) oxidize guanosines in DNA to form 8-oxo-7,8-dihydro-2-deoxyguanosine (8-oxodG), a biomarker for oxidative stress. Herein we describe a novel 64-microwell electrochemiluminescent (ECL) array enabling sensitive multiplexed detection of 8-oxodG in dsDNA without hydrolysis. Films of Nafion and reduced graphene oxide containing ECL dye [Os(bpy)(2)(phen-benz-COOH)](2+) (OsNG, {bpy= 2,2'-bipyridine and phen-benz-COOH = (4-(1,10-phenanthrolin-6-yl)benzoic acid)}) were assembled into microwells on a pyrolytic graphite wafer to detect 8-oxodG in oligonucleotides by electrochemiluminescence (ECL). DNA oxidation by Fenton's reagent or by ROS formation during redox cycles involving NADPH, Cu-II, and model metabolites was monitored. UPLC-MS/MS of oxidized DNA samples were used for calibration. Detection limit for the fluidic arrays was one 8-oxodG per 670 intact nucleobases, or 0.15%. The method is sensitive enough to evaluate DNA oxidation from biologically relevant ROS-generating reactions of Cull, NADPH, and model metabolites.
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Publisher
American Chemical Society (ACS)
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Attribution-NonCommercial-NoDerivs 3.0 Ireland