title: Chemokine Detection Using Receptors Immobilized on an SPR Sensor Surface.
abstract: Chemokines and their receptors take part in many physiological and pathological processes, and their dysregulated expression is linked to chronic inflammatory and autoimmune diseases, immunodeficiencies, and cancer. The chemokine receptors, members of the G protein-coupled receptor family, are integral membrane proteins, with seven-transmembrane domains that bind the chemokines and transmit signals through GTP-binding proteins. Many assays used to study the structure, conformation, or activation mechanism of these receptors are based on ligand-binding measurement, as are techniques to detect new agonists and antagonists that modulate chemokine function. Such methods require labeling of the chemokine and/or its receptor, which can alter their binding characteristics. Surface plasmon resonance (SPR) is a powerful technique for analysis of the interaction between immobilized receptors and ligands in solution, in real time, and without labeling. SPR measurements nonetheless require expression and purification steps that can alter the conformation, stability, and function of the chemokine and/or the chemokine receptor. In this review, we focus on distinct methods to immobilize chemokine receptors on the surface of an optical biosensor. We expose the advantages and disadvantages of different protocols used and describe in detail the method to retain viral particles as receptor carriers that can be used for SPR determinations.
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Madrid, Spain.
authors.forename: José Miguel
authors.initials: JM
authors.lastname: Rodríguez-Frade
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Madrid, Spain.
authors.forename: Laura
authors.initials: L
authors.lastname: Martínez-Muñoz
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Madrid, Spain.
authors.forename: Ricardo
authors.initials: R
authors.lastname: Villares
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Madrid, Spain.
authors.forename: Graciela
authors.initials: G
authors.lastname: Cascio
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Madrid, Spain.
authors.forename: Pilar
authors.initials: P
authors.lastname: Lucas
authors.affiliation: Department of Cell Biology, Universidad Complutense de Madrid, Madrid, Spain.
authors.forename: Rosa P
authors.initials: RP
authors.lastname: Gomariz
authors.affiliation: Department of Immunology and Oncology, Centro Nacional de Biotecnología (CNB/CSIC), Madrid, Spain. Electronic address: mmellado@cnb.csic.es.
authors.forename: Mario
authors.initials: M
authors.lastname: Mellado
mesh_terms: D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods; D018925:Chemokines / Q000032:analysis*; D006801:Humans; D055767:Immobilized Proteins / Q000737:chemistry* / Q000378:metabolism; D019707:Receptors, Chemokine / Q000737:chemistry* / Q000378:metabolism; D043562:Receptors, G-Protein-Coupled / Q000737:chemistry / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000295:instrumentation* / Q000379:methods; D014771:Virion / Q000737:chemistry* / Q000378:metabolism
chemical_list: D018925:Chemokines; D055767:Immobilized Proteins; D019707:Receptors, Chemokine; D043562:Receptors, G-Protein-Coupled
keywords: Chemokine receptors; Chemokines; Surface plasmon resonance; Viral particles