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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.
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.
Keyword(s): electrochemiluminescence; oxidative stress; DNA oxidation sensor; microwell array; [os(bpy)(2)(phen-benz-cooh)](2+); tandem mass-spectrometry; DNA-damage; liquid-chromatography; electrogenerated chemiluminescence; electron-transfer; stress; cancer; carcinogenesis; mechanisms; biomarker
Publication Date:
Type: Journal article
Peer-Reviewed: Unknown
Institution: NUI Galway
Publisher(s): American Chemical Society (ACS)
First Indexed: 2019-03-23 06:49:03 Last Updated: 2019-03-23 06:49:03