title: A New Surface Plasmon Resonance Assay for In Vitro Screening of Mannose-Binding Lectin Inhibitors.
abstract: Mannose-binding lectin (MBL) is a circulating protein that acts as a soluble pattern recognition molecule of the innate immunity. It binds to carbohydrate patterns on the surface of pathogens or of altered self-cells, with activation of the lectin pathway of the complement system. Recent evidence indicates that MBL contributes to the pathophysiology of ischemia-reperfusion injury and other conditions. Thus, MBL inhibitors offer promising therapeutic strategies, since they prevent the interaction of MBL with its target sugar arrays. We developed and characterized a novel assay based on surface plasmon resonance for in vitro screening of these compounds, which may be useful before the more expensive and time-consuming in vivo studies. The assay measures the inhibitor's ability to interfere with the binding of murine MBL-A or MBL-C, or of human recombinant MBL, to mannose residues immobilized on the sensor chip surface. We have applied the assay to measure the IC50 of synthetic glycodendrimers, two of them with neuroprotective properties in animal models of MBL-mediated injuries.
authors.affiliation: Department of Molecular Biochemistry and Pharmacology, IRCSS-Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
authors.forename: Matteo
authors.initials: M
authors.lastname: Stravalaci
authors.affiliation: Department of Neuroscience, IRCSS-Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy Department of Anesthesia and Critical Care Medicine, Fondazione IRCCS Ca'Granda-Ospedale Maggiore Policlinico, Milan, Italy.
authors.forename: Daiana
authors.initials: D
authors.lastname: De Blasio
authors.affiliation: Department of Neuroscience, IRCSS-Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
authors.forename: Franca
authors.initials: F
authors.lastname: Orsini
authors.affiliation: Department of Neuroscience, IRCSS-Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Perego
authors.affiliation: Department of Chemistry, University of Milan, Milan, Italy.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Palmioli
authors.affiliation: Department of Chemistry, University of Milan, Milan, Italy.
authors.forename: Giulio
authors.initials: G
authors.lastname: Goti
authors.affiliation: Department of Chemistry, University of Milan, Milan, Italy.
authors.forename: Anna
authors.initials: A
authors.lastname: Bernardi
authors.affiliation: Department of Neuroscience, IRCSS-Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy.
authors.forename: Maria-Grazia
authors.initials: MG
authors.lastname: De Simoni
authors.affiliation: Department of Molecular Biochemistry and Pharmacology, IRCSS-Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy marco.gobbi@marionegri.it.
authors.forename: Marco
authors.initials: M
authors.lastname: Gobbi
mesh_terms: D000818:Animals; D002241:Carbohydrates / Q000737:chemistry / Q000302:isolation & purification*; D050091:Dendrimers / Q000737:chemistry / Q000627:therapeutic use; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D007113:Immunity, Innate / Q000235:genetics; D037601:Mannose-Binding Lectin / Q000037:antagonists & inhibitors* / Q000737:chemistry; D051379:Mice; D015427:Reperfusion Injury / Q000188:drug therapy / Q000378:metabolism; D020349:Surface Plasmon Resonance / Q000379:methods*
chemical_list: D002241:Carbohydrates; D050091:Dendrimers; D037601:Mannose-Binding Lectin
keywords: glycodendrimers; immunoassay; inhibitor screening; mannose binding lectin; surface plasmon resonance