GenWay/CEA/GWB-9E60F0/1x96 Assays

Description:

ProductName:CEA

#ofSamples:1x96Assays

IntendedUse:ImmunoenzymaticcolorimetricmethodforquantitativedeterminationofCEAinhumanserum.

Introduction:Carcinoembryonicantigen(CEA)isaglycoprotein,withamolecularweightof180kDa,involvedincelladhesion.Itisnormallyproducedduringfetaldevelopment,buttheproductionofCEAstopsbeforebirth.Therefore,itisnotusuallypresentinthebloodofhealthyadults,althoughlevelsareraisedinheavysmokers.CEAwasidentifiedinhumancoloncancertissueextracts.ItwaslaterfoundthatserumfromindividualswithcolorectalandothercarcinomashadhigherlevelsofCEAthanhealthyindividualsandcanbeusedtomonitortheresponsetocoloncancertreatment.
CEAandrelatedgenesmakeuptheCEAfamilybelongingtotheimmunoglobulinsuperfamily.
ThemostfrequentcancerwhichcausesanincreasedCEAiscancerofthecolonandrectum.Othersincludecancersofthepancreas,stomach,breast,lung,andcertaintypesofthyroidandovariancancer.BenignconditionswhichcanelevateCEAincludesmoking,infections,inflammatoryboweldisease,pancreatitis,cirrhosisoftheliver,andsomebenigntumorsinthesameorgansinwhichanelevatedCEAindicatescancer.ChemotherapyandrADIationtherapycancauseatemporaryriseinCEAduetothedeathoftumorcellsandreleaseofCEAintothebloodstream.Benigntumoursdonotusuallycauseanincreaseabove10ng/ml.

Principlesoftheassay:Inthismethod,CEAstandards,patientspecimensand/orcontrolscontainingthenativeantigenarefirstaddedtostreptavidincoatedwells.Biotinylatedmonoclonalandhorseradishperoxidase(HRP)labeledantibodiesareaddedandthereactantsaremixed.ThedifferenttypesofantibodiesusedhavehighaffinityandspecificityandaredirectedagainstdistinctanddifferentepitopesofCEA.ReactionbetweenthevariousCEAantibodiesandnativeCEAoccursinthemicrowellswithoutcompetitionorstericshindranceformingasolublesandwichcomplex.
Theinteractionisillustratedbythefollowingequation:
Ka
E-Ab+AgCEA+BtnAb(m)E-Ab-AgCEA-BtnAb(m)
K-a
BtnAb(m)ByotinilatedMonoclonalAntibody(ExcessQuantity)
AgCEANativeAntigen(VariableQuantity)
E-AbenzymelabeledAntibody(ExcessQuantity)
HRP-Ab(p)-AgCEA-BtnAb(m)Antigen-AntibodiesSandwichComplex
KaRateConstantofAssociation
K-aRateConstantofDissociation

Simultaneously,thecomplexisfixedtothewellthroughthehighaffinityreactionofstreptavidinandbiotinylatedantibody.Thisinteractionisillustratedbelow:
E-Ab-AgCEA-BtnAb(m)+StreptavidinCWImmobilizedComplex
StreptavidinCWStreptavidinimmobolizedonwell.
ImmobilizedComplexAntibodies-Antigensandwichbound.
Afterequilibriumisattained,theantibody-boundfractionisseparatedfromunboundantigenbyaspiration.ThenativeantigenconcentrationisdirectlyproportionaltotheHRPactivityintheantibody-boundfraction.TheactivityoftheconjugatedHRPisquantitatedbyreactionwithTMBsubstratetoproducebluecolour.Thereactionisterminatedbyaddingstopsolutionwhichturnsthebluecolourintoyellow.Theabsorbanceismeasuredonaplatereader

StorageandStABIlity:Theclosedreagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°Cinthedark.
Openedreagentsarestablefor60dayswhenstoredat2?8°C.

LimitationsoftheTest:Bacterialcontaminationorrepeatedfreeze-thawcyclesofthespecimenmayaffecttheabsorbancevalues.
CEAhasalowclinicalsensitivityandspecificityasatumourMarker.ClinicallyanelevatedCEAvaluealoneisnotofdiagnosticvalueasatestforcancerandshouldonlybeusedinconjunctionwithotherclinicalmanifestations(observations)anddiagnosticparameters.TherearepatientswithcolorectalcancerthatdonotexhibitelevatedCEAvaluesandelevatedCEAvaluesdonotalwayschangewithprogressionorregressionofdisease.Smokersdemonstrateahigherrangeofbaselinevaluesthannon-smokers.

ReferencesGoldP,FreedmanSO,JExpMed,121,439(1965)ZamcheckN,AdvInternMed,19,413(1974)RayncaoG,ChuTM,JAMA,220,381(1972)WildD,TheImmunoassayHandbook.,StocktonPress(1994)p444SorokinJJ,etalJAMA228:49-53(1974)MackayAM,etalBr.Med.Jr.4:382-385(1974)SikorskaH,etalCancerDetectionPrev12:321-355(1988)MintonJP,etalCancer42:1422-27(1978)StaabHJ,etalAm.J.Surgery136:322-327(1978)ThomasP,etalBiochemBiophysActa1032:177-189(1990)YamashitaK,etalCancerResearch47:3451-3459(1987)HammerstromS,etalCancerResearch49:4852-58(1989)Ann.Inter.Med.1981;94:407-409

AdditionalInformation:

Name	CEA
RelatedProductNames	CEAELISA
MolecularWeight	0.5
Storage	Theclosedreagentsarestableuptotheexpirydatestatedonthelabelwhenstoredat2...8°Cinthedark. Openedreagentsarestablefor60dayswhenstoredat2?8°C.
IntendedUse	ResearchUseOnly
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