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1、J.Phys.Chem.C2007,111,8443-84508443NewAntibodyImmobilizationStrategyBasedonGoldNanoparticlesandAzureI/Multi-WalledCarbonNanotubeCompositeMembranesforanAmperometricEnzymeImmunosensorNaLi,RuoYuan,*YaqinChai,ShihongChen,HaizhenAn,andWenjuanLiChongqingKeyLaboratoryofA
2、nalyticalChemistry,CollegeofChemistryandChemicalEngineering,SouthwestUniVersity,Chongqing400715,ChinaReceiVed:December14,2006;InFinalForm:April9,2007AconvenientandeffectivestrategybasedontheuniquecharacteristicsofAzureI/multi-walledcarbonnanotube(AzureI/MWNT)compo
3、sitemembranesandtheamplificationresponseofenzymestotheantigen-antibodyreactionwasusedtodevelopahighlysensitiveamperometricimmunosensorforthedetectionofR-fetoprotein(AFP).AnoveltypeofAzureI/MWNTcompositemembranewasfabricatedbytwosteps:first,acid-pretreatedMWNTwasmo
4、difiedonthesurfaceofglassycarbonelectrode.Then,AzureIwascoatedonMWNTtoobtainanAzureI/MWNTcompositemembrane.Subsequently,goldnanoparticleswereadsorbedontotheAzureI/MWNTcompositesurfacebyelectrostaticinteractionsbetweenthenegativelychargedgoldnanoparticleandtheposit
5、ivelychargedAzureI.Later,AFPantibodieswereassembledontothesurfaceofgoldnanoparticles.Finally,horseradishperoxidase(HRP)insteadofbovineserumalbuminwasemployedtoblocksitesagainstnonspecificbindingandamplifiedthecurrentsignaloftheantigen-antibodyreaction.Severaltechn
6、iques,includingcyclicvoltammetry,UV-visabsorptionspectroscopy,scanningelectronmicroscopy,transmissionelectronmicroscopy,atomicforcemicroscopy,Ramanspectroscopy,andX-rayphotoelectronspectroscopy,havebeenemployedtocharacterizetheassemblyprocess.Becauseoftheelectroca
7、talyticabilityofHRPandMWNTforthereductionofH2O2andtheuniquepropertiesofgoldnanoparticles,andespeciallythesynergisticaugmentationofAzureIandMWNTtofacilitateelectron-transferprocesses,theimmunosensordisplayedahighsensitivity,fastanalyticaltime,andbroaderlinearrespon
8、setoAFPintworangesfrom0.1to8.0and8.0to250.0ng/mLwitharelativelylowdetectionlimitof0.04ng/mLat3timesthebackgroundnoise.Moreover,thestudiedimmunosensorpos
