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Electrochemiluminescent Array to Detect Oxidative Damage in ds-DNA Using [Os(bpy)2(phen-benz-COOH)]2+/Nafion/Graphene Films
- Source :
- ACS Sensors. 1:272-278
- Publication Year :
- 2016
- Publisher :
- American Chemical Society (ACS), 2016.
-
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.
- Subjects :
- 0301 basic medicine
Bioengineering
010402 general chemistry
Photochemistry
microwell array
01 natural sciences
Redox
Article
law.invention
stress
03 medical and health sciences
chemistry.chemical_compound
electrochemiluminescence
law
Nafion
DNA oxidation sensor
liquid-chromatography
oxidative stress
tandem mass-spectrometry
cancer
Electrochemiluminescence
electrogenerated chemiluminescence
Instrumentation
Benzoic acid
Fluid Flow and Transfer Processes
mechanisms
DNA-damage
Graphene
Oligonucleotide
Process Chemistry and Technology
DNA oxidation
electron-transfer
0104 chemical sciences
030104 developmental biology
chemistry
Reagent
biomarker
[os(bpy)(2)(phen-benz-cooh)](2+)
carcinogenesis
Subjects
Details
- ISSN :
- 23793694
- Volume :
- 1
- Database :
- OpenAIRE
- Journal :
- ACS Sensors
- Accession number :
- edsair.doi.dedup.....9dda152c421989d85084f3bbc991c1ca