title: Effects of the Surface Densities of Glycoclusters on the Determination of Their IC50 and Kd Value Determination by Using a Microarray.
abstract: Pseudomonas aeruginosa (PA) is an opportunistic bacterium involved in 10-30% of nosocomial diseases. It causes severe lung injury to cystic fibrosis patients, often leading to patient death. PA strains are multidrug resistant, thus making the design of new therapeutics a challenge for public health. One promising therapeutic option is to design glycoclusters that target the virulence factor of PA. LecA is a galactose-specific lectin that might be involved in adhesion and biofilm formation by PA. The DNA-directed immobilization (DDI) microarray is a powerful tool for screening and understanding of structure-activity relationships between glycoclusters and lectins. High-throughput and multiplexed analysis of lectin-glycocluster interactions on a DDI microarray allows measurement of IC50 and dissociation constant (Kd ) values with minute amounts of material. In order to study the robustness of the DDI microarray in determination of IC50 and Kd values, the impact of glycocluster surface density was investigated. The data obtained show that measured IC50 values were influenced by glycocluster surface density: as the density of glycoclusters increases, the measured IC50 values increase too. In contrast, the measured Kd values were not affected by glycocluster surface density, provided that the experimental conditions allow interaction between glycocluster and lectin at single-molecule level (no surface cluster effect).
authors.affiliation: Institut des Nanotechnologies de Lyon (INL), UMR 5270 CNRS, Ecole Centrale de Lyon, Université de Lyon, 36, Avenue Guy de Collongue, 69134, Ecully, France.
authors.forename: Lucie
authors.initials: L
authors.lastname: Dupin
authors.affiliation: Institut des Nanotechnologies de Lyon (INL), UMR 5270 CNRS, Ecole Centrale de Lyon, Université de Lyon, 36, Avenue Guy de Collongue, 69134, Ecully, France.
authors.forename: Francesca
authors.initials: F
authors.lastname: Zuttion
authors.affiliation: Institut des Nanotechnologies de Lyon (INL), UMR 5270 CNRS, Ecole Centrale de Lyon, Université de Lyon, 36, Avenue Guy de Collongue, 69134, Ecully, France.
authors.forename: Thomas
authors.initials: T
authors.lastname: Géhin
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR 5247 CNRS, UM, ENSCM, Université de Montpellier, Place E. Bataillon, CC1704, 34095, Montpellier Cedex 5, France.
authors.forename: Albert
authors.initials: A
authors.lastname: Meyer
authors.affiliation: Institut des Nanotechnologies de Lyon (INL), UMR 5270 CNRS, Ecole Centrale de Lyon, Université de Lyon, 36, Avenue Guy de Collongue, 69134, Ecully, France.
authors.forename: Magali
authors.initials: M
authors.lastname: Phaner-Goutorbe
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR 5247 CNRS, UM, ENSCM, Université de Montpellier, Place E. Bataillon, CC1704, 34095, Montpellier Cedex 5, France.
authors.forename: Jean-Jacques
authors.initials: JJ
authors.lastname: Vasseur
authors.affiliation: Institut des Nanotechnologies de Lyon (INL), UMR 5270 CNRS, Ecole Centrale de Lyon, Université de Lyon, 36, Avenue Guy de Collongue, 69134, Ecully, France.
authors.forename: Eliane
authors.initials: E
authors.lastname: Souteyrand
authors.affiliation: Institut des Biomolécules Max Mousseron (IBMM), UMR 5247 CNRS, UM, ENSCM, Université de Montpellier, Place E. Bataillon, CC1704, 34095, Montpellier Cedex 5, France. morvan@univ-montp2.fr.
authors.forename: François
authors.initials: F
authors.lastname: Morvan
authors.affiliation: Institut des Nanotechnologies de Lyon (INL), UMR 5270 CNRS, Ecole Centrale de Lyon, Université de Lyon, 36, Avenue Guy de Collongue, 69134, Ecully, France. yann.chevolot@ec-lyon.fr.
authors.forename: Yann
authors.initials: Y
authors.lastname: Chevolot
mesh_terms: D018829:Adhesins, Bacterial / Q000737:chemistry / Q000378:metabolism*; D001422:Bacterial Adhesion; D018441:Biofilms; D031541:Fluorescence Resonance Energy Transfer; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism*; D020128:Inhibitory Concentration 50; D007700:Kinetics; D046228:Microarray Analysis; D018625:Microscopy, Atomic Force; D011485:Protein Binding; D011550:Pseudomonas aeruginosa / Q000235:genetics / Q000378:metabolism*; D037521:Virulence Factors
chemical_list: D018829:Adhesins, Bacterial; D006023:Glycoproteins; C102929:LecA protein, bacteria; D037521:Virulence Factors
keywords: Pseudomonas aeruginosa; dissociation constants; glycoconjugates; lectins; microarrays; surface density