NANOZ-ONIC stands for carbon NANOtubes, NOse(Z) and electrONIC.
The project has for ambition to develop an original bioinspired approach for further design of biosensors for non-invasive, painless and real-time detection of volatile biomarkers in exhaled breath of patients. Such biomarkers have been identified for several severe diseases including different cancers, diabetes mellitus and neurodegeneration. A fast, simple and miniaturized breath-diagnostic system has obvious advantages for early detection of these pathologies, for increasing considerably the number of diagnosed persons and for point-of-care monitoring.
While electronic noses (e-noses) already exist for some pathologies like asthma, diagnosing diseases from breath analysis is generally impeded by the lack of one or several technical requirements (sensibility, selectivity, simplicity, versatility…), the ideal technology being analogous to the natural olfactory system.
To meet the challenge of a bio-inspired, two technologies were interfaced in this project : the Ion Channel-Coupled Receptors (ICCR) and the single-walled Carbon NanoTube-Field Effect Transistors (swCNT-FET). ICCR are original biosensors created in the IBS laboratory by linking G Protein Coupled Receptors (GPCRs) to an ion channel. Recognition of compounds by the receptor is transduced into electrical signal by the ion channel. A library of ICCRs with natural olfactory receptors would provide a coveted olfactory map of samples for accurate and multiplexed diagnostics. To detect the electrical signal with extreme sensitivities and in nanoscaled format, ICCRs are integrated in swCNT-FET transistors. The obstacles of ICCR purification, lipid bilayer insertion and stability is overcome by nanovesicles preparation. The detection of odorant with ICCR-swCNT-FET device by project members demonstrated the feasibility of this approach.
This project is a partnership between two beneficiaries : Centre National de la Recherche Scientifique (CNRS) and Seoul National University (SNU) and three laboratories : Dr. Moreau, Prof. Park and Prof. Hong.
The work performed during the project was simultaneously focused on three technical aspects : 1) development of receptor-based biosensors for detecting volatile compounds ; 2) optimization of the biological material for interfacing the biosensors with carbon nanotube Field-effect transistors (FETs) ; and 3) improvements of FETs with higher sensitivity, reusability and a surface appropriate to the analysis of gaseous samples.
The results led to the publication of 26 articles and a book chapter and the presentation of the project and results in two joint symposia between the European Federation of Biotechnology and the Asian Federation of Biotechnology.
The main progress obtained to date are :
1) in biosensor engineering :
– The development of a second generation of hybrid receptor/ion channels biosensors with enhanced signal amplitude (Garcia Fernandez et al. Sci Rep. 2021).
– The functional characterization in Xenopus oocytes of optimised human olfactory receptor provided by the laboratory of Professor Matsunami (Duke University, NC, USA) and a fish olfactory receptor.
– The electrophysiological detection of odours with olfactory receptors from other species, widening the diversity of recognized ligands.
– extracellular domains of taste receptors were also successfully used as biosensors demonstrating the feasibility of this approach (Jeong et al. ACS Appl. Mater. Interfaces 2022).
2) in production of biological materials :
– An additional and simpler method to the initial method of nanovesicles preparation from membrane of mammalian cells were developed. Thus, olfactory receptors were produced in bacteria. For their integration in FETs, they were solubilized and stabilized either in nanodiscs (Yang et al. ACS Nano 2017 ; Lee et al. Sci. Rep. 2018, Lee et al. Biosens. Bioelectron. 2020) or in dual detergent micelles (Shin et al. Sci. Rep. 2020 and Yoo et al. Sens. Actuators B Chem. 2022) for detecting odorants or toxic gas.
3) in upgrading of FETs
– A magnetic biochip was developed to improve the speed and sensitivity of detection (low non-specific binding) and to facilitate the regeneration of the surface (Yoo et al. Nanotechnology 2018).
– A sol-gel matrix was developed on the FET surface to allow the detection of volatile compound (Kim et al. Sci. Rep. 2018)
– The addition of engineered floating electrodes enable the specific, quantitative and high sensitivity detection of compounds (Pham Ba et al. ACS Appl. Mater. Interfaces 2018).
– Hybrid surfaces with carbon nanotube and gold reduced noises and resistivity (Shin et al. Adv. Electron. Mater. 2020).
1) Nanoscale hybrid systems based on carbon nanotubes for biological sensing and control. Youngtak Cho, Narae Shin, Daesan Kim, Jae Yeol Park, Seunghun Hong. Bioscience Reports. 2017 : http://europepmc.org/articles/PMC5483890
2) Magnetically-focusing biochip structures for high-speed active biosensing with improved selectivity. Haneul Yoo, Dong Jun Lee, Daesan Kim, Juhun Park, Xing Chen, Seunghun Hong. Nanotechnology. 2018 : http://stacks.iop.org/0957-4484/29/i=26/a=265501?key=crossref.dd40246d6989f553aa032a566086ad5b
3) Nanodisc-Based Bioelectronic Nose Using Olfactory Receptor Produced in Escherichia coli for the Assessment of the Death-Associated Odor Cadaverine. Heehong Yang, Daesan Kim, Jeongsu Kim, Dongseok Moon, Hyun Seok Song, Minju Lee, Seunghun Hong, Tai Hyun Park. ACS Nano. 2016 : https://hal.archives-ouvertes.fr/hal-02388831
4) Quantitative electrophysiological monitoring of anti–histamine drug effects on live cells via reusable sensor platforms. Viet Anh Pham Ba, Dong-guk Cho, Daesan Kim, Haneul Yoo, Van-Thao Ta, Seunghun Hong. Biosensors and Bioelectronics. 2017 : http://linkinghub.elsevier.com/retrieve/pii/S0956566317302294
5) Human-like smelling of a rose scent using an olfactory receptor nanodisc-based bioelectronic nose. Minju Lee, Heehong Yang, Daesan Kim, Myungjae Yang, Tai Hyun Park, Seunghun Hong. Scientific Reports. 2018 : https://www-nature-com.insb.bib.cnrs.fr/articles/s41598-018-32155-1
6) Artificial Rod and Cone Photoreceptors with Human-Like Spectral Sensitivities. Byeongho Park, Heehong Yang, Tai Hwan Ha, Hyun Seo Park, Seung Ja Oh, Yong-Sang Ryu, Youngho Cho, Hyo-Suk Kim, Juyeong Oh, Dong Kyu Lee, Chulki Kim, Taikjin Lee, Minah Seo, Jaebin Choi, Young Min Jhon, Deok Ha Woo, Seok Lee, Seok Hwan Kim, Hyuk-Jae Lee, Seong Chan Jun, Hyun Seok Song, Tai Hyun Park, Jae Hun Kim. Advanced Materials. 2018 : https://hal.archives-ouvertes.fr/hal-02187743
7) Nafion-Radical Hybrid Films on Carbon Nanotube Transistors for Monitoring Antipsychotic Drug Effects on Stimulated Dopamine Release. Viet Anh Pham Ba, Dong-guk Cho, Seunghun Hong. ACS Applied Materials & Interfaces. 2019 : https://hal.archives-ouvertes.fr/hal-02187852v1
8) Dye-functionalized Sol-gel Matrix on Carbon Nanotubes for Refreshable and Flexible Gas Sensors. Jeongsu Kim, Haneul Yoo, Viet Anh Pham Ba, Narae Shin, Seunghun Hong. Scientific Reports. 2018 : http://europepmc.org/articles/PMC6086896
9) High-Speed Lateral Flow Strategy for a Fast Biosensing with an Improved Selectivity and Binding Affinity. Dong Guk Cho, Haneul Yoo, Haein Lee, Yeol Kyo Choi, Minju Lee, Dong June Ahn, Seunghun Hong. Sensors. 2018 : https://doaj.org/toc/1424-8220
10) Bioelectronic Nose Using Olfactory Receptor-Embedded Nanodiscs. Heehong Yang, Minju Lee, Daesan Kim, Seunghun Hong, Tai Hyun Park. Olfactory Receptors (book chapter) 2018 : https://link.springer.com/protocol/10.1007/978-1-4939-8609-5_18
11) Modified Floating Electrode-Based Sensors for the Quantitative Monitoring of Drug Effects on Cytokine Levels Related with Inflammatory Bowel Diseases. Viet Anh Pham Ba, Yoo Min Han, Youngtak Cho, Taewan Kim, Byung Yang Lee, Joo Sung Kim, Seunghun Hong. ACS Applied Materials & Interfaces. 2018 : https://zenodo.org/record/3501799
12) Nanoscale mapping of noise-source-controlled hopping and tunneling conduction in domains of reduced graphene oxide. Shashank Shekhar, Hyungwoo Lee, Duckhyung Cho, Myungjae Yang, Minju Lee, Seunghun Hong. Carbon. 2019 : https://hal.archives-ouvertes.fr/hal-02384186
13) Bioelectronic sensor mimicking the human neuroendocrine system for the detection of hypothalamic-pituitary-adrenal axis hormones in human blood. Seung Hwan Lee, Minju Lee, Heehong Yang, Youngtak Cho, Seunghun Hong, Tai Hyun Park. Biosensors and Bioelectronics. 2020 : https://www.sciencedirect.com/science/article/pii/S0956566320300683?via%3Dihub
14) Semiconducting Carbon Nanotubes Embedded in a Metallic Film for a Thermistor Device with an Adjustable Temperature Coefficient and Reduced Noise Source Activities. Narae Shin, Youngtak Cho, Jeongsu Kim, Seunghun Hong. Advanced Electronic Materials. 2020 : https://onlinelibrary.wiley.com/doi/full/10.1002/aelm.202000073
15) Ion-Selective Carbon Nanotube Field-Effect Transistors for Monitoring Drug Effects on Nicotinic Acetylcholine Receptor Activation in Live Cells. Youngtak Cho, Viet Anh Pham Ba, Jin-Young Jeong, Yoonji Choi, Seunghun Hong. Sensors. 2020 : http://europepmc.org/articles/PMC7374424
16) The ligand-bound state of a G protein-coupled receptor stabilizes the interaction of functional cholesterol molecules. Laura Lemel, Katarzyna Nieścierowicz, M. Dolores García-Fernández, Leonardo Darré, Thierry Durroux, Marta Busnelli, Mylène Pezet, Fabrice Rébeillé, Juliette Jouhet, Bernard Mouillac, Carmen Domene, Bice Chini, Vadim Cherezov, Christophe J. Moreau. Journal of Lipid Research. 2021 : https://hal.archives-ouvertes.fr/hal-03180171
17) Reusable surface plasmon resonance biosensor chip for the detection of H1N1 influenza virus. Haneul Yoo, Junghyun Shin, Jieun Sim, Hyunmin Cho, Seunghun Hong. Biosensors and Bioelectronics. 2020 : https://www.sciencedirect.com/science/article/abs/pii/S0956566320305534?via%3Dihub
18) Micelle-stabilized Olfactory Receptors for a Bioelectronic Nose Detecting Butter Flavors in Real Fermented Alcoholic Beverages. Narae Shin, Seung Hwan Lee, Viet Anh Pham Ba, Tai Hyun Park, Seunghun Hong. Scientific Reports. 2020 : https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7270175/
19) Distinct classes of potassium channels fused to GPCRs as electrical signaling biosensors. M. Dolores García-Fernández ; Franck C. Chatelain ; Hugues Nury ; Anna Moroni ; Christophe Moreau. Cell Reports Methods. 2021 : https://hal.univ-grenoble-alpes.fr/hal-03518152
20) Mapping the nanoscale effects of charge traps on electrical transport in grain structures of indium tin oxide thin films. Hyesong Jeon ; Jeongsu Kim ; Shashank Shekhar ; Jeehye Park ; Seunghun Hong. Nanoscale Advances. 2021 : https://doi.org/10.1039/D1NA00175B
21) Olfactory receptor-based CNT-FET sensor for the detection of DMMP as a simulant of sarin. Jin Yoo ; Daesan Kim ; Heehong Yang ; Minju Lee ; So-ong Kim ; Hwi Jin Ko ; Seunghun Hong ; Tai Hyun Park. Sensors and Actuators B : Chemical. 2022 : https://doi.org/10.1016/j.snb.2021.131188
22) Reusable surface amplified nanobiosensor for the sub PFU/mL level detection of airborne virus. Junghyun Shin ; Hyeong Rae Kim ; Pan Kee Bae ; Haneul Yoo ; Jeongsu Kim ; Yoon-Ji Choi ; Aeyeon Kang ; Wan S. Yun ; Yong-Beom Shin ; Jungho Hwang ; Seunghun Hong. Scientific Reports. 2021 : https://doaj.org/article/e4963c581313496dafacb7bc6b772d38
23) Nanoscale mapping of wavelength-selective photovoltaic responses in H- and J-aggregates of azo dye-based solar cell films. Shashank Shekhar ; Inkyoung Park ; Jeongsu Kim ; Myungjae Yang ; Duckhyung Cho ; Seunghun Hong. Journal of Materials Chemistry A. 2020 : https://doi.org/10.1039/d0ta07328h
24) Dipolar Noise in Fluorinated Molecular Wires. Mingyu Jung, Shashank Shekhar, Duckhyung Cho, Myungjae Yang, Jeehye Park, Seunghun Hong. Nanomaterials. 2022 : https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9031467/
25) Ultrasensitive Bioelectronic Tongue Based on the Venus Flytrap Domain of a Human Sweet Taste Receptor. Jin-Young Jeong, Yeon Kyung Cha, Sae Ryun Ahn, Junghyun Shin, Yoonji Choi, Tai Hyun Park, Seunghun Hong. ACS Appl Mater Interfaces. 2022 : https://hal.archives-ouvertes.fr/hal-03668818
26) Evaluation of site-selective drug effects on GABA receptors using nanovesicle-carbon nanotube hybrid devices. Inkyoung Park, InwooYang, Youngtak Cho, Yoonji Choi, Junghyun Shin, Shashank Shekhar, Seung Hwan Lee, Seunghun Hong. Biosensors and Bioelectronics. 2022 : https://hal.archives-ouvertes.fr/hal-03668844
27) Elicitation of potent SARS-CoV-2 neutralizing antibody responses through immunization with a versatile adenovirus-inspired multimerization platform. Christopher Chevillard, Axelle Amen, Solène Besson, Dalil Hannani, Isabelle Bally, Valentin Dettling, Evelyne Gout, Christophe J Moreau, Marlyse Buisson, Salomé Gallet, Daphna Fenel, Emilie Vassal-Stermann, Guy Schoehn, Pascal Poignard, Marie-Claire Dagher, Pascal Fender. Molecular Therapy. 2022 : https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8828441/