Development of a Quantitative Bead Capture Assay for Soluble IL-7 Receptor Alpha in Human Plasma 英文参考文献.docVIP

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Development of a Quantitative Bead Capture Assay for Soluble IL-7 Receptor Alpha in Human Plasma 英文参考文献.doc

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Development of a Quantitative Bead Capture Assay for Soluble IL-7 Receptor Alpha in Human Plasma 英文参考文献

DevelopmentofaQuantitativeBeadCaptureAssayfor SolubleIL-7ReceptorAlphainHumanPlasma SylvieFaucher1*,AngelaM.Crawley2,3,WendyDecker1,AliceSherring1,DragicaBogdanovic1 ,Tao Ding1,MicheleBergeron1,JonathanB.Angel2,3,4,PaulSandstrom1 1NationalHIVandRetrovirologyLaboratories,PublicHealthAgencyofCanada,Ottawa,Canada,2OttawaHospitalResearchInstitute,Ottawa,Canada,3Departmentof Biochemistry,MicrobiologyandImmunology,UniversityofOttawa,Ottawa,Canada,4DivisionofInfectiousDiseases,OttawaHospital-GeneralCampus,Ottawa,Canada Abstract Background:IL-7isanessentialcytokineinT-celldevelopmentandhomeostasis.ItbindstotheIL-7Rreceptor,acomplexof theIL-7Ra(CD127)andcommonc(CD132)chains.ThereissignificantinterestinevaluatingtheexpressionofCD127on humanT-cellsasitoftendecreasedinmedicalconditionsleadingtolymphopenia.Previousreportsshowedtheusefulness ofCD127asaprognosticmarkerinviralinfectionssuchasHIV,CMV,EBVandHCV.AsolubleCD127(sCD127)isreleasedin plasmaandmaycontributetodiseasepathogenesisthroughitscontrolonIL-7activities.MeasuringsCD127isimportantto define its role and may complement existing markers used in lymphopenic disease management. We describe a new quantitativeassayforthemeasurementofsCD127inplasmaandreportsCD127concentrationsinhealthyadults. Methodology/Principal Findings: We developed a quantitative bead-based sCD127 capture assay. Polyclonal CD127- specificantibodieswerechosenforcaptureandabiotinylatedmonoclonalanti-CD127antibodywasselectedfordetection. TheassaycandetectnativesCD127andrecombinantsCD127whichservedasthecalibrator.Theanalyticalperformanceof theassaywascharacterizedandtheconcentrationandstabilityofplasmasCD127inhealthyadultswasdetermined.The assay’s range was 3.2–1000ng/mL. The concentration of plasma sCD127 was 1646104ng/mL with over a log variation betweensubjects.IndividualsCD127concentrationsremainedstablewhenmeasuredseriallyduringaperiodofuptoone year. Conclusions/Significance:ThisisthefirstreportonthequantificationofplasmasCD127inapopulationofhealthyad