title: Immobilization of rhodopsin on a self-assembled multilayer and its specific detection by electrochemical impedance spectroscopy.
abstract: Rhodopsin, the G protein-coupled receptor (GPCR) which mediates the sense of vision, was prepared from calf eyes and used as receptor enriched membrane fraction. In this study it was immobilized onto gold electrode by two different techniques: Langmuir-Blodgett (LB) and a strategy based on a self-assembled multilayer. We demonstrated that Langmuir and LB films of rhodopsin are not stable. Thus, in this study a new protein multilayer was prepared on gold electrode by building up layer-by-layer a self-assembled multilayer. It is composed of a mixed self-assembled monolayer formed by MHDA and biotinyl-PE, followed by a biotin-avidin system which allows binding of biotinylated antibody specific to rhodopsin. The immobilization of rhodopsin in membrane fraction, by the specific antibody bound previously on self-assembled multilayer, was monitored with electrochemical impedance spectroscopy (EIS). In addition, the specificity and sensitivity of this self-assembled multilayer system to the presence of rhodopsin were investigated. No effect was observed when the system was in contact with olfactory receptor I7 in membrane fraction used for control measurements. All these results demonstrate that rhodopsin can be immobilized efficiently, specifically, quantitatively and stably on gold electrode through the self-assembled multilayer.
authors.affiliation: Centre de Génie Electrique de Lyon (CEGELY), Ecole Centrale de Lyon, B.P.163, 69134 Ecully Cedex, France.
authors.forename: Yanxia
authors.initials: Y
authors.lastname: Hou
authors.forename: Salwa
authors.initials: S
authors.lastname: Helali
authors.forename: Aidong
authors.initials: A
authors.lastname: Zhang
authors.forename: Nicole
authors.initials: N
authors.lastname: Jaffrezic-Renault
authors.forename: Claude
authors.initials: C
authors.lastname: Martelet
authors.forename: Jasmina
authors.initials: J
authors.lastname: Minic
authors.forename: Tatiana
authors.initials: T
authors.lastname: Gorojankina
authors.forename: Marie-Annick
authors.initials: MA
authors.lastname: Persuy
authors.forename: Edith
authors.initials: E
authors.lastname: Pajot-Augy
authors.forename: Roland
authors.initials: R
authors.lastname: Salesse
authors.forename: Francois
authors.initials: F
authors.lastname: Bessueille
authors.forename: Josep
authors.initials: J
authors.lastname: Samitier
authors.forename: Abdelhamid
authors.initials: A
authors.lastname: Errachid
authors.forename: Vladimir
authors.initials: V
authors.lastname: Akimov
authors.forename: Lino
authors.initials: L
authors.lastname: Reggiani
authors.forename: Cecilia
authors.initials: C
authors.lastname: Pennetta
authors.forename: Eleonora
authors.initials: E
authors.lastname: Alfinito
mesh_terms: D000327:Adsorption; D015374:Biosensing Techniques / Q000295:instrumentation / Q000379:methods*; D020099:Coated Materials, Biocompatible / Q000032:analysis* / Q000737:chemistry*; D003460:Crystallization / Q000379:methods; D017097:Electric Impedance; D004563:Electrochemistry / Q000379:methods*; D008567:Membranes, Artificial; D011485:Protein Binding; D012243:Rhodopsin / Q000032:analysis* / Q000737:chemistry* / Q000648:ultrastructure; D013057:Spectrum Analysis / Q000379:methods*; D013499:Surface Properties
chemical_list: D020099:Coated Materials, Biocompatible; D008567:Membranes, Artificial; D012243:Rhodopsin