Human sensory-mimicking systems, such as electronic brains, tongues, skin, and ears, have been promoted for use in improving social welfare. However, no significant achievements have been made in mimicking the human nose due to the complexity of olfactory sensory neurons. Combinational coding of human olfactory receptors (hORs) is essential for odorant discrimination in mixtures, and the development of hOR-combined multiplexed systems has progressed slowly. Here, we report the first demonstration of an artificial multiplexed superbioelectronic nose (MSB-nose) that mimics the human olfactory sensory system, leading to high-performance odorant discriminatory ability in mixtures. Specifically, portable MSB-noses were constructed using highly uniform graphene micropatterns (GMs) that were conjugated with two different hORs, which were employed as transducers in a liquid-ion gated field-effect transistor (FET). Field-induced signals from the MSB-nose were monitored and provided high sensitivity and selectivity toward target odorants (minimum detectable level: 0.1 fM). More importantly, the potential of the MSB-nose as a tool to encode hOR combinations was demonstrated using principal component analysis.
Cadaverine (CV), a death-associated odor, is an important target molecule for various sensor applications, including the evaluation of food spoilage. In this study, we developed an oriented nanodisc (ND)-functionalized bioelectronic nose (ONBN), based on carbon nanotube transistors and nanodiscs embedded with an olfactory receptor produced in Escherichia coli (E. coli) for detection of CV. To fabricate ONBN devices, a trace-amine-associated receptor 13c (TAAR13c) binding to CV was produced in E. coli, purified, reconstituted into NDs, and assembled, in the desired orientation, onto a carbon- nanotube-based field-effect transistor with floating electrodes. The ONBN showed high performance in terms of sensitivity and selectivity. Moreover, the ONBN was used to measure CV in diverse real-food samples for the determination of food freshness. These results indicate ONBN devices can be utilized to evaluate the quality of food samples quantitatively, which should enable versatile practical applications such as food safety and preservative development. Moreover, the ONBN could provide a useful tool for detection of corpses, which could be practically used in disaster responses.
In the brain and central nervous system, dopamine plays a crucial role as a neurotransmitter or a local chemical messenger for interneuronal communication. Dopamine is associated with renal, hormonal, and cardiovascular systems. Additionally, dopamine dysfunction is known to cause serious illnesses, such as Parkinson's disease and Alzheimer's disease. Therefore, dopamine detection is essential for medical diagnosis and disease prevention and requires a novel strategy with high sensitivity and selectivity and a rapid response. Herein, we present a novel human dopamine receptor (hDRD1)-conjugated multidimensional conducting polymer nanofiber (NF) membrane for the selective and sensitive detection of dopamine. The membrane, which consists of multidimensional carboxylated poly(3,4-ethylenedioxythiophene) (MCPEDOT) NFs with nanorods, is used as a transistor in a liquid-ion gated field-effect transistor (FET)-based biosensor. Interestingly, hDRD1 is first expressed in Escherichia coli before it is immobilized onto the MCPEDOT NF. The hDRD1-MCPEDOT NF-based FET exhibits a rapid real-time response (<2 s) with high dopamine selectivity and sensitivity performance (approximately 100 fM). Furthermore, this FET device can be integrated into a poly(dimethylsiloxane)-based microfluidic system and also can retain its high performance in the integrated system, which results in the generation of large-scale dopamine biosensors with a novel geometry.
Field-effect transistor (FET) devices based on conductive nanomaterials have been used to develop biosensors. However, development of FET-based biosensors that allow efficient stability, especially in the gas phase, for obtaining reliable and reproducible responses remains a challenge. In this study, we developed a nanodisc (ND)-functionalized bioelectronic nose (NBN) based on a nickel (Ni)-decorated carboxylated polypyrrole nanoparticle (cPPyNP)-FET that offers the detection of liquid and gaseous cadaverine (CV). The TAAR13c, specifically binding to CV, which is an indicator of food spoilage, was successfully constructed in NDs. The NBN was fabricated by the oriented assembly of TAAR13c-embedded NDs (T13NDs) onto the transistor with Ni/cPPyNPs. The NBN showed high performance in selectivity and sensitivity for the detection of CV, with excellent stability in both aqueous and gas phases. Moreover, the NBN allowed efficient measurement of corrupted real-food samples. It demonstrates the ND-based device can allow the practical biosensor that provides high stability in the gas phase.
Olfactory receptors (ORs), belonging to the Gprotein coupled receptor (GPCR) family, are very difficult to be overexpressed, purified and reconstituted because of their hydrophobicity and complicated structure. These receptors bind to their specific ligands, thus their specificity is very useful for application as a bioelectronic nose. Furthermore, highly purified and well-reconstituted human olfactory receptor (hOR) can be used in various fields, such as in protein-interaction research, drug screening, and analysis of the hOR structure. In this study, human olfactory receptor, hOR2AG1, was produced with high purity and functionally reconstituted in detergent micelles. The hOR2AG1 was overexpressed in Escherichia coli (E. coli) with glutathione S-transferase (GST) and 6xHis-tag as an inclusion body. The hOR2AG1 fusion protein was solubilized in buffer containing sodium dodecyl sulfate (SDS) and purified using Ni-NTA chromatography. The GST domain was removed using proteolytic cleavage before elution from the column. After purification, the hOR2AG1 was successfully reconstituted using nonionic detergents and methyl-β-cyclodextrin. Finally highly purified and well-reconstituted hOR was obtained, and its biological characteristics were confirmed by using circular dichroism (CD) spectrum and tryptophan fluorescence assay. These results can be applied to develop protein-based sensing systems including a bioelectronic nose and to analyze the native hOR structure using solid-state NMR, X-ray crystallography, or neutron scattering.
We report a strategy for the human-like smelling of a rose scent utilizing olfactory receptor nanodisc (ND)-based bioelectronic nose devices. In this strategy, a floating electrode (FE)-based carbon nanotube (CNT) field effect transistor (FET) was functionalized with human olfactory receptor 1A2 (hOR1A2)-embedded NDs (hOR1A2NDs). The hOR1A2NDs responded to rose scent molecules specifically, which were monitored electrically using the underlying CNT-FET. This strategy allowed us to quantitatively assess the contents of geraniol and citronellol, the main components of a rose scent, as low as 1 fM and 10 fM, respectively. In addition, it enabled us to selectively discriminate a specific rose odorant from other odorants. Significantly, we also demonstrated that the responses of hOR1A2NDs to a rose scent could be strongly enhanced by enhancer materials like a human nose. Furthermore, the method provided a means to quantitatively evaluate rose scent components in real samples such as rose oil. Since our method allows one to quantitatively evaluate general rose scent ingredients just like a human nose, it could be a powerful strategy for versatile basic research and various applications such as fragrance development.
The field-effect transistor (FET) has been used in the development of diagnostic tools for several decades, leading to high-performance biosensors. Therefore, the FET platform can provide the foundation for the next generation of analytical methods. A major role of G-protein-coupled receptors (GPCRs) is in the transfer of external signals into the cell and promoting human body functions; thus, their principle application is in the screening of new drugs. The research community uses efficient systems to screen potential GPCR drugs; nevertheless, the need to develop GPCR-conjugated analytical devices remains for next-generation new drug screening. In this study, we proposed an approach for studying receptor agonism and antagonism by combining the roles of FETs and GPCRs in a dopamine receptor D1 (DRD1)-conjugated FET system, which is a suitable substitute for conventional cell-based receptor assays. DRD1 was reconstituted and purified to mimic native binding pockets that have highly discriminative interactions with DRD1 agonists/antagonists. The real-time responses from the DRD1-nanohybrid FET were highly sensitive and selective for dopamine agonists/antagonists, and their maximal response levels were clearly different depending on their DRD1 affinities. Moreover, the equilibrium constants (K) were estimated by fitting the response levels. Each K value indicates the variation in the affinity between DRD1 and the agonists/antagonists; a greater K value corresponds to a stronger DRD1 affinity in agonism, whereas a lower K value in antagonism indicates a stronger dopamine-blocking effect.
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