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1、DevelopmentofELISAMehodsandKitsforRapidDetectionandanColloidalGoldImmunochromategraphicSripforDetectionofSulfonamidesAbstract4-aminobenzoate(PBPA)acetylamino-benzenesulfonylchloride(ASC)asrawmaterial,afternucleophilicsubstitution,esterhydrolysisreactionsyntheticsulfadrugsmothernucleusstructurebenzo
2、icaminobenzenesulfonamide(SH),heapplicationofmassspectrometry(ESI-MS)andproton(1nuclearmagneticresonancespectroscopyH-NMR)identificationofSH.Massspectrometryresults-showthatthe[M]=291.03766withthetheoreticalvalueofthe[M]=292match,indicatingthesuccess1ofSHsynthesis;NMR(H-NMR)datamatchthestructureofS
3、Hcompound.Usingthemixedanhydridemethod,thelaboratorysynthesisofsulphuraminemedicinemothernucleusstructure(SH)wasconnetedwithovalbumin(OVA)andbovineserumalbumin(BSA),syntheticcoatingantigenOVA-SHandimmunogenBSA-SH.Ultravioletscanning(UV)andSDS-PAGEmethodswereestablished.ImmuneBalb/CmicebyBSA-SH,indi
4、rectELISAandindirectcompetitiveELISAmethodforscreeningofcellfusionratbycellfusionstandby;hybridomatechnologytoestablishtheSHcellline.InBalb/Cmice’vivoinducedascitespreparedSHMab.TheresultsshowedthatBSA-SHartificialantigenwassynthesizedsuccessfully,SHandBSAconjugationratioofabout11.6:1.5G1,5G2and8D2
5、3strainssensitivetospecifichybridomacelllinewas23successfullyscreened.Theircelltiterofthesupernatantsolutionwere1:6.40×10,1:1.28×10and25551:3.20×10,ascitestiterwere1:2.56×10,1:5.12×10and1.28×10.TheMabof5G2showedgoodsensitivitywithanIC50of82ng/mLtoSH.ThecrossreactiveratesoftheMabafterpurifiedtoSulfa
6、monomethoxine(SMM),Sulfametoxydiazine(SMD),Sulfamerazine(SM),Sulfadimethoxine(SDM),Sulfacetamide(SCT),Sulfaclozinesodiummonohydrate(SCZ),Sulfathiazole(ST),Sulfamethoxazole(SMZ),Sulfaphenazole(SPH),Sulfamethazine(SM2),Sulfasulfonamide(SN)andSulfaquinoxaline(SQ)were108%,86%,105%,85.6%,72%,26%,32%,54.
7、6%,43%,4.6%,0.5%and76%.SHMabofhigh-titer,sensitivityandspecificityhasbeengenerated.Theeggalbumin(OVA)andthenucleusofsulfanilamideartificialhapten(SH)bymixedanhydridemethod,connectingthefeedratioof1:2ofthepa