ELISA方法类型及原理

2025/07/23TypesandFundamentalsofELISATechniqueReporter:CONTENTSCatalogue01TypesofELISAMethod02BasicPrinciplesofELISA03ApplicationScenariosofDifferentELISAMethodsTypesofELISAMethod01DirectELISADirectELISADirectELISAemploysthecoatingofamicroplatewithanantigen,subsequenttowhichanenzyme-taggedantibody,capableofdirectlyconnectingwiththeantigen,isadded.TheprocessismorestraightforwardandinvolvesfewerstepsthanotherformsofELISA.IndirectELISAIndirectELISAMethodologyIndirectELISAdetectsantigensbyusinganenzyme-labeledsecondaryantibodythatbindstotheprimaryantibody,providingameasurablesignal.IndirectELISA'sKeyComponentsThemethodinvolvescoatingtheplatewiththeantigen,addingtheprimaryantibody,andthenasecondaryantibodyconjugatedwithanenzyme.IndirectELISATheIndirectELISAMethodologyTheprocessentailsfixingtheantigen,followedbyincubationwiththeprimaryantibody,thenthesecondaryantibody,andfinallyaddingasubstrateforcolormanifestation.ApplicationsofIndirectELISAIndirectELISAiswidelyusedinresearchfordetectingantibodiesinserum,measuringantigen-specificantibodies,andindiagnostictests.SandwichELISASandwichELISASandwichELISAemploysapairofantibodies,eachrecognizingauniqueepitopeonthetargetantigen.Oneantibodyisattachedtoasolidsurface,whiletheotheristaggedwithadetectablemarker,enablingthedetectionandmeasurementoftheantigen.CompetitiveELISACompetitiveELISAIncompetitiveELISA,there'sacompetitionoccurringbetweentheantigenwithinthesampleandanenzyme-taggedantigenintermsofbindingwithanantibody.Thehighertheamountofantigendetectedinthesample,thelesserthequantityofthelabeledantigenwillattachtotheantibody,whichconsequentlyleadstoareducedsignal.BasicPrinciplesofELISA02Antigen-AntibodySpecificBindingMasteringthefundamentalsofantigen-antibodyconnectionAntigensserveasstimulicapableofinitiatinganimmunereaction,whereasantibodiesareproteinssynthesisedbytheimmunesystemtocounteractantigens.ThedistinctivenessintheirinteractionformsthebasisofELISA.SpecificBindingofAntigenandAntibodyAntigen-antibodyinteractionisfundamentalinELISAprocedures.Enzyme-LinkedImmunosorbentAssay(ELISA)usesthisspecificbindingtodetectandquantifyantigensorantibodiesinasample,makingitcrucialtograsptheprinciplesofthisinteraction.Antigen-AbSpecificInteractionRoleofenzymesinsignalamplificationThereporterenzymelinkedtotheantibodyorantigenconvertsthebindingprocessintoadetectablesignal,enhancingitforanalysisandquantitationinanELISA.EnzymeMarkingandSubstrateInteractionCompetitiveELISAThecompetitiveELISAprocessentailsacompetitionforbindingbetweentheantigenpresentinthesampleandanenzyme-taggedantigen,alloccurringatafinitenumberofantibody-bindingsites.ELISA技术应用的多样场景03RapidScreeningELISAMethodDirectELISADirectELISAproceduresentailapplyingtheantigentothemicroplatesurface,thenintroducinganenzyme-linkedantibodywhichattachestotheantigendirectly.AntibodyDetectionviaIndirectEnzyme-LinkedImmunosorbentAssaySandwichELISASandwichELISAemploysadual-antibodysystemforantigenidentification,wheretheinitialantibodybindstotheantigenandthesubsequentantibodyconfirmsitspresence,ensuringbothhighspecificityandsensitivity.AntibodyDetectionviaIndirectEnzyme-LinkedImmunosorbentAssayDirectELISATheELISAtechnique,knownasdirectELISA,usesasingleantibodytodirectlyconnectwiththeantigen,followedbydetectionviaanenzyme-linkedantibody,renderingtheprocesssimple.AntibodyDetectionviaIndirectEnzyme-LinkedImmunosorbentAssayIndirectELISATheindirectELISAemploysaprimaryantibodyforantigenbindingandasecondaryantibodyfortheprimaryantibody'sdetection,providingversatilityandsignalamplification.AntibodyDetectionviaIndirectEnzyme-LinkedImmunosorbentAssayCompetitiveELISAELISA竞争法通过检测已知量标记抗原与抗体结合的抑制能力来测定抗原含量，适用于小分子物质。AntigenQuantificationviaSandwichELISATechniq