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Analog Integrated Circuits and Signal Processing

, Volume 57, Issue 3, pp 197–203 | Cite as

Nanobiosensors based on individual olfactory receptors

  • 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
Article

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.

Keywords

Biosensors Electronic nose Olfactory receptors Nanotechnology Odorant 

Notes

Acknowledgments

This work was financially supported by the SPOT-NOSED Project of the European Community (IST-2001-38739), the Ile-de-France region, in the framework of a SESAME contract (2002/AO1497), and the PICASSO program (HF2004-0055) funded by EGIDE, and the MIUR PRIN prot.2005091492.

SPOT-NOSED (Single PrOTein NanObioSEnsor griD array) was a project funded by the European Community for the implementation of the Fifth Framework Programme, IST priority, Future and Emerging Technologies, IST-2001-38899 (January 2003–April 2006). The Project Coordinator was Prof. J. Samitier (U. Barcelona), and the SPOT-NOSED original idea was by A. Errachid and G. Gomila (U. Barcelona).

References

  1. 1.
    Malnic, B., Hirono, J., Sato, T., & Buck, L. B. (1999). Combinatorial receptor codes for odors. Cell, 96, 713–723.CrossRefGoogle Scholar
  2. 2.
    Gomila, G., Casuso, I., Errachid, A., Ruiz, O., Pajot, E., Minic, J., Gorojankina, T., Persuy, M. A., Aioun, J., Salesse, R., Bausells, J., Villanueva, G., Rius, G., Hou, Y., Jaffrezic, N., Pennetta, C., Alfinito, E., Akimov, V., Reggiani, L., Ferrari, G., Fumagalli, L., Sampietro, M., & Samitier, J. (2006). Artificial nose integrating biological olfactory receptors and NEMS. Sensors and Actuators B, 116, 66–71.CrossRefGoogle Scholar
  3. 3.
    Minic, J., Persuy, M. A., Godel, E., Aioun, J., Connerton, I., Salesse, R., & Pajot-Augy, E. (2005). Functional expression of olfactory receptors in yeast and development of a bioassay for odorant screening. The FEBS Journal, 272, 524–537.CrossRefGoogle Scholar
  4. 4.
    Single PrOTein NanObioSEnsor griD array IST-2001–38899-SPOT-NOSED European project (2005). E. Pajot & European Consortium, 15th IUPAB & 5th EBSA International Biophysics Congress, Montpellier, France, August 27–September 1.Google Scholar
  5. 5.
    Gomila, G., Errachid, A., Bessueille, F., Ruiz, O., Casuso, I., Pajot-Augy, E., Minic, J., Gorojankina, T., Persuy, M.-A., Salesse, R., Villanueva, G., Bausells, J., Pennetta, C., Alfinito, E., Della Sala, F., Akimov, V., Reggiani, L., Hou, Y., Jaffrezic, N., Ferrari, G., Fumagalli, L., Sampietro, M., & Samitier, J. (2005). Production, immobilization and electrical characterization of olfactory receptors for olfactory nanobiosensor development, Euronanoforum 2005, Edinburgh, Scotland, September 5–9.Google Scholar
  6. 6.
    Casuso, I., Pla-Roca, M., Gomila, G., Minic Vidic, J., Persuy, M. A., Aioun, J., Pajot-Augy, E., Salesse, R., & Samitier, J. Native nanovesicles containing olfactory receptors adsorbed onto gold electrodes for biosensor applications. An atomic force microscopy study. Material Science and Engineering C (in press).Google Scholar
  7. 7.
    Vidic, J., Pla-Roca, M., Grosclaude, J., Persuy, M. A., Monnerie, R., Errachid, A., Hou, Y., Jaffrezic-Renault, N., Salesse, R., Pajot-Augy, E., & Samitier, J. (2007). Gold surface functionalization and patterning for specific immobilization of olfactory receptors carried by nanosomes. Analytical Chemistry, 79, 3280–3290.CrossRefGoogle Scholar
  8. 8.
    Minic, J., Grosclaude, J., Aioun, J., Persuy, M. A., Gorojankina, T., Salesse, R., Pajot-Augy, E., Hou, Y., Helali, S., Jaffrezic-Renault, N., Bessueille, F., Errachid, A., Gomila, G., Ruiz, O., & Samitier, J. (2005). Immobilization of native membrane-bound rhodopsin on biosensor surfaces. Biochimica et Biophysica Acta, 1724, 324–332.Google Scholar
  9. 9.
    Hou, Y., Helali, S., Zhang, A., Jaffrezic-Renault, N., Martelet, C., Minic, J., Gorojankina, T., Persuy, M. A., Pajot-Augy, E., Salesse, R., Bessueille, F., Samitier, J., Errachid, A., Akinov, V., Reggiani, L., Pennetta, C., & Alfinito, E. (2006). Immobilization of rhodopsin on a self-assembled multilayer and its specific detection by electrochemical impedance spectroscopy. Biosensors and Bioelectronics, 21(7), 1393–1402.CrossRefGoogle Scholar
  10. 10.
    Hou, Y., Jaffrezic-Renault, N., Martelet, C., Zhang, A., Minic, J., Gorojankina, T., Persuy, M. A., Pajot-Augy, E., Salesse, R., Akimov, V., Reggiani, L., Pennetta, C., Alfinito, E., Ruiz, O., Gomila, G., Samitier, J., & Errachid, A. (2007). A novel detection strategy for odorant molecules based on controlled bioengineering of rat olfactory receptor I7 (OR I7). Biosensors and Bioelectronics, 22, 1550–1555.CrossRefGoogle Scholar
  11. 11.
    Rodriguez Segui, S., Pla, M., Minic, J., Pajot-Augy, E., Salesse, R., Hou, Y., Jaffrezic-Renault, N., Mills, C. A., Samitier, J., & Errachid, A. (2006). Detection of olfactory receptor I7 self assembled multilayer formation and immobilization using a quartz crystal microbalance. Analytical Letters, Special Issue: Second International Workshop on “Biosensors for Food Safety and Environmental Monitoring” Agadir, Morocco, November 2005 (Vol. 39, pp. 1735–1745).Google Scholar
  12. 12.
    Minic Vidic, J., Grosclaude, J., Persuy, M. A., Aioun, J., Salesse, R., & Pajot-Augy, E. (2006) Quantitative assessment of olfactory receptors activity in immobilized nanosomes: A novel concept for bioelectronic nose. Lab on a Chip, 6, 1026–1032.CrossRefGoogle Scholar
  13. 13.
    Pennetta, C., Akimov, V., Alfinito, E., Reggiani, L., Gorojankina, T., Minic, J., Pajot-Augy, E., Persuy, M. A., Salesse, R., Casuso, I., Errachid, A., Gomila, G., Ruiz, O., Samitier, J., Hou, Y., Jaffrezic, N., Ferrari, G., Fumagalli, L., & Sampietro, M. (2006). Towards the realization of nanobiosensors based on G protein-coupled receptors. In C. S. S. R. Kumar (Ed.), Nanodevices for the life sciences (series: Nanotechnologies for the life sciences) (Vol. 4, pp. 217–240) Wiley-VCH.Google Scholar
  14. 14.
    Benilova, I. V., Minic Vidic, J., Pajot-Augy, E., Soldatkin, A. P., Martelet, C. & Jaffrezic-Renault, N. Electrochemical study of human olfactory receptor OR 17–40 stimulation by odorants in solution. Material Science and Engineering C (in press).Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2007

Authors and Affiliations

  • Vladimir Akimov
    • 1
  • Eleonora Alfinito
    • 1
  • Joan Bausells
    • 2
  • Iryna Benilova
    • 3
  • Ignacio Casuso Paramo
    • 4
  • Abdelhamid Errachid
    • 4
  • Giorgio Ferrari
    • 5
  • Laura Fumagalli
    • 5
  • Gabriel Gomila
    • 4
  • Jeanne Grosclaude
    • 6
  • Yanxia Hou
    • 7
  • Nicole Jaffrezic-Renault
    • 8
  • Claude Martelet
    • 3
  • Edith Pajot-Augy
    • 9
  • Cecilia Pennetta
    • 1
  • Marie-Annick Persuy
    • 9
  • Mateu Pla-Roca
    • 4
  • Lino Reggiani
    • 1
  • Santiago Rodriguez-Segui
    • 4
  • Oscar Ruiz
    • 4
  • Roland Salesse
    • 9
  • Josep Samitier
    • 4
  • Marco Sampietro
    • 5
  • Alexey P. Soldatkin
    • 10
  • Jasmina Vidic
    • 6
  • Guillermo Villanueva
    • 2
  1. 1.Dipartimento di Ingegneria dell’Innovazione NNL-LNFMUniversita’ del SalentoLecceItaly
  2. 2.Centro Nacional de Microelectronica (IMB-CSIC)BellaterraSpain
  3. 3.Laboratoire de Sciences AnalytiquesUniversité Claude Bernard - Lyon 1Villeurbanne CedexFrance
  4. 4.Labarotori de Nanobioenginyeria-CREBEC, Parc Cientific de Barcelona and Department d’ElectronicaUniversitat de BarcelonaBarcelonaSpain
  5. 5.Dipartimento di Elettronica ed InformazionePolitecnico di MilanoMilanoItaly
  6. 6.INRA, Unité de Virologie et Immunologie MoléculairesJouy-en-Josas CedexFrance
  7. 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)GrenobleFrance
  8. 8.Centre de Génie Electrique de Lyon (CEGELY)Ecully CedexFrance
  9. 9.INRA, Neurobiologie de l’Olfaction et de la Prise AlimentaireEquipe Récepteurs et Communication ChimiqueJouy-en-Josas CedexFrance
  10. 10.Institute of Molecular Biology and Genetics National Academy of Sciences of UkraineKyivUkraine

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