Nanobiosensors based on individual olfactory receptors
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.
Pajot-Augy, E INRA, Equipe Recepteurs & Commun Chim, F-78352 Jouy En Josas, France 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 Politecn Milan, 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
Cited References Count:14
Record created on 2013-08-06, modified on 2016-08-09