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Title

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

AuthorsVega, Beatriz; Martínez Muñoz, Laura; Holgado, Borja L.; Lucas, Pilar; Rodríguez Frade, José M.; Calle Martín, Ana ; Rodríguez-Fernández, José Luis ; Lechuga, Laura M. ; Rodríguez, José F.; Gutiérrez-Gallego, Ricardo; Mellado, Mario
Issue DateAug-2011
PublisherSociety for Leukocyte Biology
CitationJournal of Leukocyte Biology 90(2): 399-408 (2011)
AbstractUse of SPR-based biosensors is an established method for measuring molecular interactions. Their application to the study of GPCRs is nonetheless limited to detergent-solubilized receptors that can then be reconstituted into a lipid environment. Using the chemokine receptor CXCR4 and its specific ligand CXCL12, we outline here a highly reproducible biosensor method based on receptor presentation on the surface of lentiviral particles; the approach is simple and does not require the use of antibodies to achieve correct receptor orientation on the sensorchip surface. We measured the kinetic parameters of CXCR4/CXCL12 binding in a single step and in real time and evaluated the effect of GAG presentation of chemokines on this interaction. The data indicate that at low concentrations, soluble heparin modulates CXCR4/CXCL12 interaction and at high concentrations, abrogates binding. These observations suggest that in addition to their known role in modulating local chemokine availability, GAG affect the receptor/ligand interaction, although their influence on affinity parameters is very limited. The method will also be useful for quantifying these biomarkers in biological fluids and for the development of high-throughput screening for their antagonists.
Publisher version (URL)http://dx.doi.org/10.1189/jlb.1010565
URIhttp://hdl.handle.net/10261/51115
DOI10.1189/jlb.1010565
Identifiersissn: 0741-5400
e-issn: 1938-3673
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