V. Akimov, E. Alfinito, J. Bausells, I. Benilova, I.C. Paramo, A. Errachid, G. Ferrari, L. Fumagalli, G. Gomila, J. Grosclaude, Y.X. Hou, N. Jaffrezic-Renault, C. Martelet, E. Pajot-Augy, C. Pennetta, M.A. Persuy, M. Pla-Roca, L. Reggiani, S. Rodriguez-Segui, S. Akimov, M. Sampietro, V. Alfinito, E. Bausells, J. Benilova, I.V. Casuso, I. Errachid, A. Ferrari, G. Fumagalli, L. Gomila, G. Grosclaude, J. Hou, Y. Jaffrezic-Renault, N. Martelet, C. Pajot-Augy, E. Pennetta, C. Persuy, M.A. Pla-Roca
INRA, Equipe Recepteurs & Commun Chim, F-78352 Jouy En Josas, France; Univ. Salento, Dipartimento Ingn Innovaz NNL LNFM, I-73100 Lecce, Italy; CSIC, Ctr Nacl Microelect IMB, Bellaterra 08193, Spain; Univ. Lyon 1, CNRS, Sci Analyt Lab, UMR 5180, F-69622 Villeurbanne, France; Univ. Barcelona, Lab Nanobioengn CREBEC, Barcelona 108028, Spain; Univ. Barcelona, Dept Elect, Barcelona 108028, Spain; Politecnico Milano, Dipartimento Elettron & Informaz, I-20133 Milan, Italy; INRA, Unite Virol & Immunol Mol, F-78352 Jouy En Josas, France; CEA, Dept. Microtechnol Biol & Sante DTBS, Lab Elect & Technol Informat LETI, F-38054 Grenoble, France; Ecole Cent Lyon, Ctr Genie Elect Lyon CEGELY, F-69134 Ecully, France; Natl Acad Sci Ukraine, Inst Mol Biol & Genet, UA-03143 Kiev, Ukraine
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.
Analog integrated circuits and signal processing 57 , pp. 197–203
IMMOBILIZATION, NANOSOMES, RHODOPSIN
Reggiani Lino, Pennetta Cecilia, Sampietro Marco, Ferrari Gabriele
IFN – Istituto di fotonica e nanotecnologie, INFM – Centro di responsabilità scientifica INFM
External IDs
CNR OAI-PMH: oai:it.cnr:prodotti:167366