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1、Immunoassayusingprobe-labellingimmunogoldnanoparticleswithsilverstainingenhancementviasurface-enhancedRamanscatteringShupingXu,aXiaohuiJi,bWeiqingXu,*aXiaolingLi,aLianyingWang,bYubaiBai,*bBingZhaoaandYukihiroOzakicaKeyLaboratoryforSupramolecularStructureandMaterialofMinistryofEduc
2、ation,JilinUniversity,Changchun130021,P.R.ChinabCollegeofChemistry,JilinUniversity,Changchun130021,P.R.China.E-mail:wqxu@jlu.edu.cn;Fax:+86-431-8980729;Tel:+86-431-8498912cDepartmentofChemistry,SchoolofScienceandTechnology,Kwansei-GakuinUniversity,Sanda,Hyogo669-1337,JapanReceived
3、17thOctober2003,Accepted1stDecember2003FirstpublishedasanAdvanceArticleontheweb11thDecember2003Thispaperreportsanovelimmunoassaybasedonsurface-enhancedRamanscattering(SERS)andimmunogoldlabellingwithsilverstainingenhancement.ImmunoreactionsbetweenimmunogoldcolloidsmodifiedbyaRaman-
4、activeprobemolecule(e.g.,4-mercaptobenzoicacid)andantigens,whichwerecapturedbyantibody-assembledchipssuchassiliconorquartz,weredetectedviaSERSsignalsofRaman-activeprobemolecule.Alltheself-assembledstepsweresubjectedtothemeasurementsofultraviolet-visible(UV-vis)spectratomonitorthef
5、ormationofasandwichstructureontoasubstrate.Theimmunoassaywasperformedbyasandwichstructureconsistingofthreelayers.ThefirstlayerwascomposedofimmobilizedantibodymoleculesofmousepolyclonalantibodyagainstHepatitisBvirussurfaceantigen(PAb)onasiliconorquartzsubstrate.Thesecondlayerwasthe
6、complementaryHepatitisBvirussurfaceantigen(Antigen)moleculescapturedbyPAbonthesubstrate.Thethirdlayerwascomposedoftheprobe-labellingimmunogoldnanoparticles,whichweremodifiedbymousemonoclonalantibodyagainstHepatitisBvirussurfaceantigen(MAb)and4-mercaptobenzoicacid(MBA)astheRaman-ac
7、tiveprobeonthesurfaceofgoldcolloids.Aftersilverstainingenhancement,theantigenisidentifiedbyaSERSspectrumofMBA.AworkingcurveoftheintensityofaSERSsignalat1585cm21duetothen8aaromaticringvibrationofMBAversustheconcentrationofanalyte(Antigen)wasobtainedandthenon-optimizeddetectionlimit
8、fortheHepatitisBvirussurfaceantig
