Nanobiosensors based on individual olfactory receptors |
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Authors: | Vladimir Akimov Eleonora Alfinito Joan Bausells Iryna Benilova Ignacio Casuso Paramo Abdelhamid Errachid Giorgio Ferrari Laura Fumagalli Gabriel Gomila Jeanne Grosclaude Yanxia Hou Nicole Jaffrezic-Renault Claude Martelet Edith Pajot-Augy Cecilia Pennetta Marie-Annick Persuy Mateu Pla-Roca Lino Reggiani Santiago Rodriguez-Segui Oscar Ruiz Roland Salesse Josep Samitier Marco Sampietro Alexey P. Soldatkin Jasmina Vidic Guillermo Villanueva |
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Affiliation: | 1. Dipartimento di Ingegneria dell’Innovazione NNL-LNFM, Universita’ del Salento, Via Arnesano, Lecce, 73100, Italy 2. Centro Nacional de Microelectronica (IMB-CSIC), Campus UAB, Bellaterra, 08193, Spain 3. Laboratoire de Sciences Analytiques, Université Claude Bernard - Lyon 1, UMR-CNRS 5180, Villeurbanne Cedex, 69622, France 4. Labarotori de Nanobioenginyeria-CREBEC, Parc Cientific de Barcelona and Department d’Electronica, Universitat de Barcelona, C/Marti i Franqués, Barcelona, 108028, Spain 5. Dipartimento di Elettronica ed Informazione, Politecnico di Milano, Pz. Leonardo da Vinci 32, Milano, 20133, Italy 6. INRA, Unité de Virologie et Immunologie Moléculaires, Domaine de Vilvert, Jouy-en-Josas Cedex, 78352, France 7. Département des Microtechnologies Pour la Biologie et la Santé (DTBS), Laboratoire d’électronique et de technologies de l’information (LETI), Commissariat à l’Energie Atomique (CEA), 17 Rue des Martyrs, Grenoble, 38054, France 8. Centre de Génie Electrique de Lyon (CEGELY), Ecole Centrale de Lyon, Ecully Cedex, 69134, France 9. INRA, Neurobiologie de l’Olfaction et de la Prise Alimentaire, Equipe Récepteurs et Communication Chimique, Domaine de Vilvert, Jouy-en-Josas Cedex, 78352, France 10. Institute of Molecular Biology and Genetics, National Academy of Sciences of Ukraine, 150 Zabolotnogo str., Kyiv, 03143, Ukraine
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Abstract: | The animal olfactory system represents the gold standard of biosensors, due to its ability to identify and discriminate thousands of odorant compounds with very low thresholds. Using olfactory receptors (ORs) as sensing elements instead of chemical sensors, biosensors would benefit the naturally optimized molecular recognition of odorants to develop a new generation of bioelectronic noses. The purpose of SPOT-NOSED European project was the development of nanobiosensors based on single ORs anchored between nanoelectrodes, to mimic the performances of natural olfactory system. Nanobiosensors arrays could then increase odorant sensitivity or widen the odorant detection spectrum. ORs were expressed in yeasts plasmic membrane, and their functionality tested in whole yeasts. Then, nanosomes bearing the ORs were prepared from S. cerevisiae, and Surface Plasmon Resonance was performed on nanosomes for quantitative evaluation of OR response to odorant stimulation. ORs retain full activity and discrimination power in immobilized nanosomes, thus allowing their use in the fabrication of the nanobiosensors. Nanoelectrodes were fabricated using conventional photolithography and focused ion beam milling, with sizes in adequation with the nanosomes. ORs borne by nanosomes were specifically immobilized onto conducting substrates via mixed Self Assembled Monolayers, neutravidin and specific antibody to the ORs. The process was optimized by microcontact printing, and the anchored nanovesicles visualized by Atomic Force Microscopy. A transimpedance preamplifier suited for low-noise wide-bandwidth measurements was designed to be directly connected to the nanoelectrodes. Electrochemical Impedancemetric Spectroscopy detected significant changes upon electrodes functionalization, grafting of ORs carried by nanosomes, and ORs conformational change induced by odorant binding. |
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