表面等离子体子共振传感器研究头孢菌素类药物与白蛋白的相互作用

1211111#1211300122410082AOTF-SPRSPRHSAAOTF1.17×103L/mol44.22%2.55×102L/mol14.79%AOTF-SPR2005-08-152005-11-21No.2003183035No.200405611HSA12～4SPR5～7AOTFAOTFSPR89AOTF-SPR-22.1N-N-hydroxysuccinimideNHSN--N-1-Ethyl-3-3-dimethylaminopropylcarbodiimideEDCHSAMPASigmapH7.40.01mol/LPBS2.2AOTF-SPR110AOTF440～790nmAOTFPSSPRPP50nm180µLPMT3420067FENXIHUAXUEChineseJournalofAnalyticalChemistry7967～9701AOTF-SPRFig.1Schematicdiagramofthewavelengthmodulationacousto-optictunalbefilter-surfaceplasmonresonanceAOTF-SPRbiosensorAOTFAOTF0～5V790～440nmAOTFλ=817.6094-130.9617E-18.1799E2+26.945E3-8.3720E4+0.93962E5+0.00177E6-0.00474E7r=0.9998SPRSPRΔEEoESPRΔEE-Eo2.3HSAPBSAOTFHSA2.42.4.1DPMD+PMkak/0dPMD11dΔE/dt=kaDΔEmax-ΔE-kdΔ1DkakdΔEmaxPMDΔEtdΔE/dt=0ΔEΔEΔEeqΔEeq1ΔEeq/D=KAΔEmax-KAΔEeq2KA=ka/kdΔEeq/DΔEeqKA2.4.2ΔEeqPBSSPRdΔE/dt=-kdΔE3lnΔE/ΔE0=-kdt-t04t0kdKA=ka/kdKAkd24kaSPRKA#12KA=#DtPt-#Dt×Dt-#Dt5PtPt=0.68mmol/LDtDt=0.01mmol/L33.1PBS10mmol/LMPAMPA86934SPR7minSPR95%MPA-0.024VPBSMPA-0.023V2.10nmPBSMPA&&SAuMPA40g/LEDCNHSEDCMPANHSMPAHSAMPANHSSPRPBS2HSAMPAFig.2KineticcurveofhumanserumalbuminHSAonthemercaptopropionicacidMPAmonolayer-0.027V2.37nm3.2pHpH=7.4PBS5g/L2HSAMPA90min45minSPRHSA24hPBS-0.040V3.58nm3.3HSA31mmol/LHSASPRFig.3SPRspectrumof1mmol/LcefaclorA.phosphatebuttersolutionB.1mmol/LcefaclorHSAMPAAOTFSPRHSA31mmol/LHSASPRHSA4HSA14abFig.4Relationshipsbetweentheconcentrationforacefaclorbceftriaxoneandtheshiftofresonantvoltage96971Table1Drug/serumalbuminequilibriumdissociationconstantsdeterminedbySPRCompoundkaL/molS-1kdS-1KAL/molx%x14%Cefaclor1.03×1028.81x10-21.17×10344.222.5Ceftriaxone1.16×1014.54×10-22.55×10214.79References1PeterTJ.AllaboutalbulminBiochemistryGeneticsandMedicalApplications.NewYorkAcademicPress.19962XuYanHuangHanguoShenHanxi.ChineseJ.Anal.Chem.199826121494～14973PangYHYangLLShaoMSDongCThompsonM.JournalofPhotochemistryandPhotobiologyBBiology200580139～1444AscenziPBocediANotariSMenegattiEFasanoM.BiochemicalandBiophysicalResearchCommunications2005334481～4865HomolaJ.Anal.Bioanal.Chem.2003377528～5396CacciatoreGPetzMRachidSHakenbeckRBergwerffAA.Anal.Chim.Acta2004520105～1157AhmadARamakrishnanAMcLeanMABreauAP.BiosensorsandBioelectronics200318399～4048JoryMJBradberryGWCannPSSamblesJR.Meas.Sci.Technol.199561193～12009CarusoaFJoryMJBradberryGWCannPSSamblesJRFurlongbDN.J.Appl.Phys.1998831023～102810TianYZhaoLWSongDQLiuXCaoYBPengZHLiuZYZhangHQ.Anal.Chim.Acta200451197～10411OberRJCavesJWardES.Anal.Biochem.200331257～6512RichRLDayYSNMortonTAMyszkaDG.Anal.Biochem.2001296197～20713GengHongyeWangShaohua.Practicalpharmacology.BeijingPeople'sMedicalPublishingHouse200371～72StudyofInteractionofCephalosporinswithHumanSerumAlbuminbyAcousto-opticTunableFilter-SurfacePlasmonResonanceBiosensorTianYuan12ChenYanhua1BiShuyun1LiuXia1SongDaqian1SunYing1ZhangHanqi#121CollegeofChemistryJilinUniversityChangchun1300122StateKeyLaboratoryofChemo/BiosensingandChemometricsHunanUninversityHunanProvinceChangsha410082AbstractAnacousto-optictunablefilter-surfaceplasmonresonanceAOTF-SPRbiosensorbasedonwave-length-modulationwasappliedtothestudyoftheinteractionofcephalosporinswithhumanserumalbuminHSA.HSAwasimmobilizedbyastandardaminecouplingmethod.ByfastvoltagescanningofAOTFtheprocessesofbiosensor'ssurfaceself-assemblingandinteractionofcephalosporinswithhumanserumalbuminweremonitoredinrealtime.Withaknownconcentrationofthecephalosporinsaffinitiesandbindingpercent-agesofthecephalosporinsforhumanserumalbuminHSAwereobtained.ForcefaclorKAis1.17×103L/molandthebindingpercentagesis44.22%ForceftriaxoneKAis2.55×102L/molandthebindingpercentagesis14.79%.TheresultsindicatethatAOTF-SPRbiosensorcanbeappliedtothedetectionofinteractionofdrugswithserumalbumin.KeywordsAcousto-optictunablefiltersurfaceplasmonresonancebiosensorceftriaxonecefaclorhumanserumalbuminReceived15August2005accepted21November200507934