Journal
ACS SENSORS
Volume 1, Issue 3, Pages 272-278Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acssensors.5b00189
Keywords
electrochemiluminescence; oxidative stress; DNA oxidation sensor; microwell array; [Os(bpy)(2)(phen-benz-COOH)](2+)
Funding
- National Institute of Environmental Health Sciences (NIEHS), NIH [ES03154]
- Science Foundation Ireland [12/TIDA/B2382]
- Science Foundation Ireland (SFI) [12/TIDA/B2382] Funding Source: Science Foundation Ireland (SFI)
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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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