Technical Advance: Surface plasmon resonance‐based analysis of CXCL12 binding using immobilized lentiviral particles

Journal of Leukocyte Biology: An Official Publication of the Reticuloendothelial Society

关键词: chemokine;    biosensor;    glycosaminoglycan;    affinity constant;    chemokine receptor;   

DOI  :  10.1189/jlb.1010565

学科分类：生理学

来源: Federation of American Societies for Experimental Biology

【 摘 要 】

UseofSPR‐basedbiosensorsisanestablishedmethodformeasuringmolecularinteractions.TheirapplicationtothestudyofGPCRsisnonethelesslimitedtodetergent‐solubilizedreceptorsthatcanthenbereconstitutedintoalipidenvironment.UsingthechemokinereceptorCXCR4anditsspecificligandCXCL12,weoutlinehereahighlyreproduciblebiosensormethodbasedonreceptorpresentationonthesurfaceoflentiviralparticles;theapproachissimpleanddoesnotrequiretheuseofantibodiestoachievecorrectreceptororientationonthesensorchipsurface.WemeasuredthekineticparametersofCXCR4/CXCL12bindinginasinglestepandinrealtimeandevaluatedtheeffectofGAGpresentationofchemokinesonthisinteraction.Thedataindicatethatatlowconcentrations,solubleheparinmodulatesCXCR4/CXCL12interactionandathighconcentrations,abrogatesbinding.Theseobservationssuggestthatinadditiontotheirknownroleinmodulatinglocalchemokineavailability,GAGaffectthereceptor/ligandinteraction,althoughtheirinfluenceonaffinityparametersisverylimited.Themethodwillalsobeusefulforquantifyingthesebiomarkersinbiologicalfluidsandforthedevelopmentofhigh‐throughputscreeningfortheirantagonists...

【 授权许可】

CC BY