• 文献标题:   Electrochemiluminescent Array to Detect Oxidative Damage in dsDNA Using [Os(bpy)(2)(phen-benz-COOH)](2+)/Nafion/Graphene Films
  • 文献类型:   Article
  • 作  者:   BIST I, SONG B, MOSA IM, KEYES TE, MARTIN A, FORSTER RJ, RUSLING JF
  • 作者关键词:   electrochemiluminescence, oxidative stres, dna oxidation sensor, microwell array, os bpy 2 phenbenzcooh 2+
  • 出版物名称:   ACS SENSORS
  • ISSN:   2379-3694
  • 通讯作者地址:   Univ Connecticut
  • 被引频次:   14
  • DOI:   10.1021/acssensors.5b00189
  • 出版年:   2016

▎ 摘  要

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.