title: Amine coupling versus biotin capture for the assessment of sulfonamide as ligands of hCA isoforms.
abstract: This work was dedicated to the development of a reliable SPR method allowing the simultaneous and quick determination of the affinity and selectivity of designed sulfonamide derivatives for hCAIX and hCAXII versus hCAII, in order to provide an efficient tool to discover drugs for anticancer therapy of solid tumors. We performed for the first time a comparison of two immobilization approaches of hCA isoforms. First one relies on the use of an amine coupling strategy, using a CM7 chip to obtain higher immobilization levels than with a CM5 chip and consequently the affinity with an higher precision (CV% < 10%). The second corresponds to a capture of proteins on a streptavidin chip, named CAP chip, after optimization of biotinylation conditions (amine versus carboxyl coupling, biotin to protein ratio). Thanks to the amine coupling approach, only hCAII and hCAXII isoforms were efficiently biotinylated to reach relevant immobilization (3000 RU and 2700 RU, respectively) to perform affinity studies. For hCAIX, despite a successful biotinylation, capture on the CAP chip was a failure. Finally, concordance between affinities obtained for the three derivatives to CAs isozymes on both chips has allowed to valid the approaches for a further screening of new derivatives.
authors.affiliation: Univ Lille, EA 7365 - GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000 Lille, France.
authors.forename: Tiphaine
authors.initials: T
authors.lastname: Rogez-Florent
authors.affiliation: Univ Lille, EA 7365 - GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000 Lille, France; Univ. Lille, ICPAL, F-59000 Lille, France.
authors.forename: Laurence
authors.initials: L
authors.lastname: Goossens
authors.affiliation: Univ. Lille, Plate-forme d'interactions moléculaires, F-59000 Lille, France; CHU Lille, Centre de biologie pathologie génétique, F-59000 Lille, France.
authors.forename: Anne-Sophie
authors.initials: AS
authors.lastname: Drucbert
authors.affiliation: Univ. Artois, Laboratoire de la barrière hémato-encéphalique (LBHE) - EA 2465, F-62307 Lens, France.
authors.forename: Sophie
authors.initials: S
authors.lastname: Duban-Deweer
authors.affiliation: Univ. Lille, ICPAL, F-59000 Lille, France.
authors.forename: Perrine
authors.initials: P
authors.lastname: Six
authors.affiliation: Univ Lille, EA 7365 - GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000 Lille, France; Univ. Lille, ICPAL, F-59000 Lille, France.
authors.forename: Patrick
authors.initials: P
authors.lastname: Depreux
authors.affiliation: Univ. Lille, Plate-forme d'interactions moléculaires, F-59000 Lille, France; CHU Lille, Centre de biologie pathologie génétique, F-59000 Lille, France.
authors.forename: Pierre-Marie
authors.initials: PM
authors.lastname: Danzé
authors.affiliation: Univ Lille, EA 7365 - GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000 Lille, France.
authors.forename: Jean-François
authors.initials: JF
authors.lastname: Goossens
authors.affiliation: Univ Lille, EA 7365 - GRITA - Groupe de Recherche sur les formes Injectables et les Technologies Associées, F-59000 Lille, France; Univ. Lille, Plate-forme d'interactions moléculaires, F-59000 Lille, France. Electronic address: catherine.foulon@univ-lille2.fr.
authors.forename: Catherine
authors.initials: C
authors.lastname: Foulon
mesh_terms: D001710:Biotin / Q000737:chemistry*; D019921:Biotinylation; D002256:Carbonic Anhydrases / Q000737:chemistry*; D004800:Enzymes, Immobilized / Q000737:chemistry*; D006801:Humans; D007527:Isoenzymes / Q000737:chemistry; D013449:Sulfonamides / Q000737:chemistry*
chemical_list: D004800:Enzymes, Immobilized; D007527:Isoenzymes; D013449:Sulfonamides; D001710:Biotin; D002256:Carbonic Anhydrases
keywords: Anticancer therapeutics; Binding; Carbonic anhydrase; Sulfonamide derivatives; Surface plasmon resonance