Detection of Olfactory Receptor I7 Self‐Assembled Multilayer Formation and Immobilization Using a Quartz Crystal Microbalance

Seguí, S. Rodríguez; Pla, M.; Minic, Jasmina; Pajot‐Augy, Edith; Salesse, Roland; Hou, Yanxia; Jaffrézic‐Renault, Nicole; Mills, Christopher A.; Samitier, J.; Errachid, Abdelhamid

doi:10.1080/00032710600714030

Cited by 11 publications

(4 citation statements)

References 18 publications

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“…Electrochemical impedance spectroscopy (EIS) is predicted to allow the direct monitoring of electrical properties changes of a single olfactory receptor, generated by its conformational change elicited by the binding of the odorant ligand . This was already demonstrated at the millimetric scale. , Finally, mass changes due to olfactory receptor−odorant complex formation can be successfully probed using quartz crystal microbalance measurements. ,, …”

supporting

confidence: 83%

“…10,11 Finally, mass changes due to olfactory receptor-odorant complex formation can be successfully probed using quartz crystal microbalance measurements. 7,12,13 Olfactory receptors OR1740 and ORI7 carried by nanosomes exhibited a fine discrimination between odorant ligands and unrelated odorants in SPR and EIS experiments, 6,11 as previously observed with a bioluminescence reporter in yeast cells 5 or by calcium response in mammalian cells. 14 This strongly supported the fact that full receptor activity is maintained in the nanosomes.…”

mentioning

confidence: 99%

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Gold Surface Functionalization and Patterning for Specific Immobilization of Olfactory Receptors Carried by Nanosomes

et al. 2007

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There is substantial interest in engineering solid supports to achieve functional immobilization of membrane receptors both for investigation of their biological function and for the development of novel biosensors. Three simple and practical strategies for immobilization of a human olfactory receptor carried by nanosomes are presented. The basis of the functionalization of solid gold surfaces is a self-assembled monolayer (SAM) containing biotinyl groups. Biotinyl groups are subsequently used to attach neutravidin and then biotinylated monoclonal antibody directed against the receptor to allow its specific grafting. Surface plasmon resonance technique is implemented for real-time monitoring of step-by-step surface functionalization and, in addition, for testing the functional response of immobilized olfactory receptors. We show that OR1740 is functional when immobilized via a tag attached to its C-terminus, but not via its N-terminus. Finally, we demonstrate that gold surfaces can be patterned by the SAMs tested using microcontact printing. AFM images of immobilized nanosomes onto a patterned surface suggest that small nanosomes flatten and fuse into larger vesicles but do not merge into a continuous layer. The whole study emphasizes the outstanding performances of the BAT/PEGAT SAM, which could be useful for developing on-a-chip sensor formats for membrane receptor investigations and use.

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confidence: 83%

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confidence: 99%

Gold Surface Functionalization and Patterning for Specific Immobilization of Olfactory Receptors Carried by Nanosomes

et al. 2007

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“…In that study, the surface was coated with a crude membrane expressing olfactory receptor protein ODR-10 extracted from E. coli then it examined its interaction with various odorant molecules, showing a liner dose-dependent response of the piezoelectronic biosensor upon membrane extraction with the natural receptor ligand diacetyl. Similar research by Segui et al, in the same year, used a membrane fraction carrying olfactory receptor protein and rat olfactory receptor I7 as a sensing element; the study provided the first step toward developing a QCM olfactory sensor [ 42 ]. For the sensor, a self-assembled multilayer composed of a mixed MHDA-biotinyl PE self-assembled monolayer and a biotin–avidin bridge system was grafted onto the sensor surface, and a receptor-specific biotinylated antibody was used to recognize a membrane fraction containing I7 receptor protein.…”

Section: Biological Recognition Elementsmentioning

confidence: 99%

Applications and Advances in Bioelectronic Noses for Odour Sensing

Choi

et al. 2018

Sensors

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A bioelectronic nose, an intelligent chemical sensor array system coupled with bio-receptors to identify gases and vapours, resembles mammalian olfaction by which many vertebrates can sniff out volatile organic compounds (VOCs) sensitively and specifically even at very low concentrations. Olfaction is undertaken by the olfactory system, which detects odorants that are inhaled through the nose where they come into contact with the olfactory epithelium containing olfactory receptors (ORs). Because of its ability to mimic biological olfaction, a bio-inspired electronic nose has been used to detect a variety of important compounds in complex environments. Recently, biosensor systems have been introduced that combine nanoelectronic technology and olfactory receptors themselves as a source of capturing elements for biosensing. In this article, we will present the latest advances in bioelectronic nose technology mimicking the olfactory system, including biological recognition elements, emerging detection systems, production and immobilization of sensing elements on sensor surface, and applications of bioelectronic noses. Furthermore, current research trends and future challenges in this field will be discussed.

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“…The efficiency of transfer and specificity of immobilization were checked with several techniques: Surface Plasmon Resonance (SPR) [7,8], Electrochemical Impedance Spectroscopy (EIS) [9,10], Quartz Crystal Microbalance (QCM) [11], and Atomic Force Microscopy (AFM) [6,7]. Fig.…”

Section: Surface Functionalization and Functional Testsmentioning

confidence: 99%

Nanobiosensors based on individual olfactory receptors

Akimov

Alfinito

Bausells

et al. 2007

Analog Integr Circ Sig Process

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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 detectionThe overall objective of the SPOT-NOSED (Single PrOTein NanObioSEnsor griD array) project was to explore the possibility to develop an olfactory nanobiosensor array based on the electrical properties of single olfactory receptors. The olfactory nanobiosensor array would integrate elementary nanobiotransducers, each of which consisting of a set of functionalised nanoelectrodes with an olfactory receptor monolayer anchored on them. The nanoelectrode would be in charge of detecting electrically any conformational or chemical change in the olfactory receptors when these receptors bind a given odorant molecule (ligand). The array structure would increase the sensitivity and specificity of the whole olfactory sensor. In order to reach the main objective of the project, advances needed to be performed in a variety of fields such as nanotechnologies, nanoelectronics, surface functionalization, expression, grafting, and electrical modelization of olfactory receptors. It required a very strong interaction within the Consortium, between research groups from very different fields, located in Spain, Italy, and France. Most results reported in the present paper were obtained within the framework of this project. Project website: 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 c...

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Detection of Olfactory Receptor I7 Self‐Assembled Multilayer Formation and Immobilization Using a Quartz Crystal Microbalance

Cited by 11 publications

References 18 publications

Gold Surface Functionalization and Patterning for Specific Immobilization of Olfactory Receptors Carried by Nanosomes

Gold Surface Functionalization and Patterning for Specific Immobilization of Olfactory Receptors Carried by Nanosomes

Applications and Advances in Bioelectronic Noses for Odour Sensing

Nanobiosensors based on individual olfactory receptors

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