Identification of Cadmium Compounds in a Solution Using Graphene-Based Sensor Array.

Rapid detection of heavy metals in solution is necessary to ensure human health and environmental protection. Some heavy-metal compounds are present in solution as compounds instead of as ions owing to their low ionization. Therefore, the development of sensor devices for the detection of heavy-metal compounds is important. In this study, as a proof of concept, we propose a sensor device using graphene and a chelating agent, which were used to develop an identification technique for three types of cadmium compounds. Pristine-graphene and two types of chelator-modified graphene-based sensors were successfully used to detect cadmium compounds at concentrations ranging from 50 to 1000 µM. The detection time was less than 5 min. The three type of graphene-based sensors responded differently to each cadmium compound, which indicates that they detected cadmium as a cadmium compound instead of as cadmium ions. Furthermore, we successfully identified cadmium compounds by operating these three types of sensors as a sensor array on the same substrate. The results indicate that sensors that focus on heavy-metal compounds instead of heavy-metal ions can be used for the detection of heavy metals in solution.

Development of an odorant sensor with a cell-free synthesized olfactory receptor and a graphene field-effect transistor.

Animals sense odorants using olfactory receptors. Many trials have been conducted to develop artificial odorant sensors using olfactory receptors. However, the development has been hindered by the difficulty in obtaining olfactory receptors. In this study, we expressed an olfactory receptor, cOR52, using a wheat germ cell-free synthesis system. The functionality of the expressed cOR52 was confirmed by ligand concentration-dependent interactions with the mini-G protein. The expressed cOR52 was immobilized on a graphene field-effect transistor. The cOR52-modified graphene field-effect transistor exhibited a ligand-specific response between 100 nM and 100 µM. This approach seems to be applicable for other olfactory receptors. Therefore, it will be possible to develop an odorant sensor equipped with various olfactory receptors by this method.

Electrical Detection of Molecular Transformations Associated with Chemical Reactions Using Graphene Devices.

This study proposes a method to electrically detect chemical reactions that involve bond changes through reactions on graphene surfaces. To achieve a highly sensitive detection, we focused on the thiol-ene reaction that combines the maleimide and thiol groups. Graphene field-effect transistors (FETs) were used to detect the binding changes of the modified molecules. Graphene has high carrier mobility and is sensitive to changes in the electronic state of its surface. Graphene has been used as a sensor to detect low-concentration targets with high sensitivity. N-(9-Acridinyl)maleimide (NAM) was chosen as the modified molecule to immobilize maleimide on graphene through π-interaction, and methanethiol (MeSH) was set as the target thiol. The modification of NAM to graphene was first confirmed by attenuated total reflection Fourier transform infrared spectroscopy, and the modification density was 0.5 ± 0.1/nm2 through cyclic voltammetry. Owing to a bond exchange, the transfer characteristics of the graphene FET shifted by 2 V to the negative direction after being exposed to MeSH at 10 parts per billion (ppb), equivalent to 0.2 ng, under ultraviolet irradiation. With 5000 ppb of acetic acid, it only shifted 0.7 V. With 1000 ppb of ethanol and 10,000 ppb of methanol, it shifted to the positive direction by 0.4 and 0.6 V, respectively. Because the nontarget molecule showed only a slight response, a thiol-ene chemical reaction was detected. The proposed method can detect the bond-change reaction using an ultralow concentration of MeSH, which indicates that at least 10 ppb (or 0.2 ng) of MeSH was detected by the graphene FET.

Electrical detection of ppb region NO2 using Mg-porphyrin-modified graphene field-effect transistors.

The trace detection of NO2 through small sensors is essential for air quality measurement and the health field; however, small sensors based on electrical devices cannot detect NO2 with the desired selectivity and quantitativity in the parts per billion (ppb) concentration region. In this study, we fabricated metalloporphyrin-modified graphene field-effect transistors (FETs). Mg-, Ni-, Cu-, and Co-porphyrins were deposited on the graphene FETs, and the transfer characteristics were measured. With the introduction of NO2 in the ppb concentration region, the FETs of pristine graphene and Ni-, Cu-, and Co-porphyrin-modified graphene showed an insufficient response, whereas the Mg-porphyrin-modified graphene exhibited large voltage shifts in the transport characteristics. This indicates that Mg-porphyrin acts as an adsorption site for NO2 molecules. An analysis of the Dirac-point voltage shifts with the introduction of NO2 indicates that the shifts were well-fitted with the Langmuir adsorption isotherm model, and the limit of detection for NO2 was found to be 0.3 ppb in N2. The relationship between the mobility and the Dirac-point voltage shift with the NO2 concentration shows that the complex of NO2 and Mg-porphyrin behaves as a point-like charge impurity. Moreover, the Mg-porphyrin-modified graphene FETs show less response to other gases (O2, H2, acetic acid, trimethylamine, methanol, and hexane), thus indicating high sensitivity for NO2 detection. Furthermore, we successfully demonstrated the quantitative detection of NO2 in air, which is near the environmental standards. In conclusion, the results of the Mg-porphyrin-modified graphene FETs enable a rapid, easy, and selective detectability.

Selective Detection of Cu2+ Ions by Immobilizing Thiacalix[4]arene on Graphene Field-Effect Transistors.

Highly accurate quantitative detection of heavy metals is essential for environmental pollution monitoring and health safety. Here, for selective detection of Cu2+ ions with high sensitivity, thiacalix[4]arene (TCA) immobilized on graphene field-effect transistors (G-FETs) are developed. Our proposed TCA-immobilized G-FETs are successfully used to detect Cu2+ ions at concentrations ranging from 1 µM to 1 mM via changes in their transfer characteristics. Moreover, the measured transfer characteristics clearly shift only when Cu2+ ions are introduced in the buffer solution despite it containing other metal ions, including those of Na+, Mg2+, Ni2+, and Cd2+; this selective detection of Cu2+ ions is attributed to the planar arrangement of TCA on graphene. Therefore, TCA-immobilized G-FETs selectively detect Cu2+ with high sensitivity.

Photosensing System Using Photosystem I and Gold Nanoparticle on Graphene Field-Effect Transistor.

In this study, a light sensor is fabricated based on photosystem I (PSI) and a graphene field-effect transistor (FET) that detects light at a high quantum yield under ambient conditions. We immobilized PSI on a micrometer-sized graphene FET using Au nanoparticles (AuNPs) and measured the I-V characteristics of the modified graphene FET before and after light irradiation. The source-drain current (Isd) increased upon illumination, exhibiting a photoresponsivity of 4.8 × 102 A W-1, and the charge neutrality point of graphene shifted by -12 mV. This system represents the first successful photosensing system based on proteins and a solution-gated graphene FET. The probable mechanism of this negative shift can be explained by the increase in negative charge carriers in graphene induced by a hole trap in the AuNP resulting from electron transfer from the AuNP to PSI. Photoresponses were only observed in the presence of two surface-active agents, n-hexyltrimethylammonium bromide and sodium dodecylbenzenesulfonate, because they caused the formation of a hydrophobic environment on the surface of the graphene. The lipid layer of these agents caused dissociation of ascorbate ions from the graphene sheet, thereby expanding the Debye screening length of the electrolyte solution. The hydrophobic environment above graphene also enhanced hole storage in the AuNP through electron transfer from the AuNP to PSI.

Graphene Surface Acoustic Wave Sensor for Simultaneous Detection of Charge and Mass.

We have combined a graphene field-effect transistor (GFET) and a surface acoustic wave (SAW) sensor on a LiTaO3 substrate to create a graphene surface acoustic wave (GSAW) sensor. When a SAW propagates in graphene, an acoustoelectric current (IA) flows between two attached electrodes. This current has unique electrical characteristics, having both positive and negative peak values with respect to the electrolyte-gate voltage (VEg) in solution. We found that IA is controlled by VEg and the amplitude of the SAW. It was also confirmed that the GSAW sensor detects changes of electrical charge in solution like conventional GFET sensors. Furthermore, the detection of amino-group-modified microbeads was performed by employing a GSAW sensor in a phthalate buffer solution at pH 4.1. The hole current peak shifted to the lower left in the IA-VEg characteristics. The left shift was caused by charge detection by the GFET and can be explained by an increase of amino groups that have positive charges at pH 4.1. In contrast, the downward shift is thought to be due to a reduction in the amplitude of the propagating SAW because of an increase in the mass loading of microbeads. This mass loading was detected by the SAW sensor. Thus, we have demonstrated that the GSAW sensor is a transducer capable of the simultaneous detection of charge and mass, which indicates that it is an attractive platform for highly sensitive and multifunctional solution sensing.

Peptide aptamer-modified single-walled carbon nanotube-based transistors for high-performance biosensors.

Biosensors employing single-walled carbon nanotube field-effect transistors (SWCNT FETs) offer ultimate sensitivity. However, besides the sensitivity, a high selectivity is critically important to distinguish the true signal from interference signals in a non-controlled environment. This work presents the first demonstration of the successful integration of a novel peptide aptamer with a liquid-gated SWCNT FET to achieve highly sensitive and specific detection of Cathepsin E (CatE), a useful prognostic biomarker for cancer diagnosis. Novel peptide aptamers that specifically recognize CatE are engineered by systemic in vitro evolution. The SWCNTs were firstly grown using the thermal chemical vapor deposition (CVD) method and then were employed as a channel to fabricate a SWCNT FET device. Next, the SWCNTs were functionalized by noncovalent immobilization of the peptide aptamer using 1-pyrenebutanoic acid succinimidyl ester (PBASE) linker. The resulting FET sensors exhibited a high selectivity (no response to bovine serum albumin and cathepsin K) and label-free detection of CatE at unprecedentedly low concentrations in both phosphate-buffered saline (2.3 pM) and human serum (0.23 nM). Our results highlight the use of peptide aptamer-modified SWCNT FET sensors as a promising platform for near-patient testing and point-of-care testing applications.

Gold-linked electrochemical immunoassay on single-walled carbon nanotube for highly sensitive detection of human chorionic gonadotropin hormone.

A new sensitive gold-linked electrochemical immunoassay (GLEIA) for the detection of the pregnancy marker human chorionic gonadotropin (hCG) has been developed using the direct electrochemical detection of Au nanoparticles. We utilized single-walled carbon nanotube (SWCNT) microelectrodes; 24 SWCNT microelectrodes were arrayed on a single Si substrate 25×30 mm² in size, for the development of a new GLEIA (SWCNT-GLEIA). This SWCNT-GLEIA provided convenient and cost-effective tests with the required antibody and antigen sample volumes as small as 2.0 µL for a group of 4 SWCNT microelectrodes. In addition, this assay also exhibited properties of high sensitivity and selectivity benefitting from the intrinsic extraordinary features of SWCNTs. Using scanning electron microscopy, we also observed Au nanoparticle-labeled antigen-antibody complexes immobilized on the surface of the SWCNT microelectrodes. The concentration of the pregnancy marker (hCG) showed a linear relationship with the current intensity obtained from differential pulse voltammetry measurements with a limit of detection (LOD) of 2.4 pg/mL (0.024 mIU/mL) hCG. This LOD is 15 times more sensitive than a previous GLEIA, which used screen-printed carbon electrodes.

Fabrication of new single-walled carbon nanotubes microelectrode for electrochemical sensors application.

In this paper, we describe two simple different ways to fabricate an array of single-walled carbon nanotubes (SWCNT) microelectrodes from SWCNT network, grown on Si substrate, through micro-fabrication process. Two kinds of material, photoresist - organic compound and sputtered SiO(2), were used as an insulator layer for these arrays of SWCNT microelectrodes. The SWCNT microelectrodes were characterized by scanning electron microscopy (SEM), Raman spectroscopy, and electrochemical measurements. The SWCNT microelectrodes with sputtered SiO(2) as an insulator exhibit some prior advances to these used photoresist layer as insulator such as much stable in harsh condition (high active organic solvents) and high current density (24.94 µA mm(-2) compared to 2.69 µA mm(-2), respectively). In addition, the well-defined geometry of SWCNT microelectrodes is not only useful for investigating kinetics of electron transfer, but also promising candidate in electrochemical sensors application.

Label-free biosensors based on aptamer-modified graphene field-effect transistors.

A label-free immunosensor based on an aptamer-modified graphene field-effect transistor (G-FET) is demonstrated. Immunoglobulin E (IgE) aptamers with an approximate height of 3 nm were successfully immobilized on a graphene surface, as confirmed by atomic force microscopy. The aptamer-modified G-FET showed selective electrical detection of IgE protein. From the dependence of the drain current variation on the IgE concentration, the dissociation constant was estimated to be 47 nM, indicating good affinity and the potential for G-FETs to be used in biological sensors.

Chemical and biological sensing applications based on graphene field-effect transistors.

Chemical and biological sensors based on graphene field-effect transistors (G-FETs) were investigated. A single-layer of graphene was prepared by mechanical cleavage of natural graphite. The G-FETs were driven by a reference-gate operating in buffer solution, and exhibited very good transport characteristics. The G-FETs detected the pH value of the solution with high precision. The Dirac point shifted in the positive direction with increasing pH of the solution. The detection limit (signal/noise=3) for measuring changes in the pH of the solution was estimated to be 0.025, indicating the high sensitivity of the G-FETs. Moreover, the devices electrically detected proteins with different charge types. The drain current decreased (increased) when positively (negatively) charged proteins were added to the solution. These results indicate that the G-FETs are among the most suitable candidates for FET-based chemical and biological sensors.

Electrolyte-gated graphene field-effect transistors for detecting pH and protein adsorption.

We investigated electrolyte-gated graphene field-effect transistors (GFETs) for electrical detecting pH and protein adsorptions. Nonfunctionalized single-layer graphene was used as a channel. GFETs immersed in an electrolyte showed transconductances 30 times higher than those in a vacuum and their conductances exhibited a direct linear increase with electrolyte pH, indicating their potential for use in pH sensor applications. We also attempted to direct surface-protein adsorption and showed that the conductance of GFETs increased with exposure to a protein at several hundred picomolar. The GFETs thus acted as highly sensitive electrical sensors for detecting pH and biomolecule concentrations.

Label-free electrical detection using carbon nanotube-based biosensors.

Label-free detections of biomolecules have attracted great attention in a lot of life science fields such as genomics, clinical diagnosis and practical pharmacy. In this article, we reviewed amperometric and potentiometric biosensors based on carbon nanotubes (CNTs). In amperometric detections, CNT-modified electrodes were used as working electrodes to significantly enhance electroactive surface area. In contrast, the potentiometric biosensors were based on aptamer-modified CNT field-effect transistors (CNTFETs). Since aptamers are artificial oligonucleotides and thus are smaller than the Debye length, proteins can be detected with high sensitivity. In this review, we discussed on the technology, characteristics and developments for commercialization in label-free CNT-based biosensors.

Label-free protein biosensor based on aptamer-modified carbon nanotube field-effect transistors.

We have fabricated label-free protein biosensors based on aptamer-modified carbon nanotube field-effect transistors (CNT-FETs) for the detection of immunoglobulin E (IgE). After the covalent immobilization of 5'-amino-modified 45-mer aptamers on the CNT channels, the electrical properties of the CNT-FETs were monitored in real time. The introduction of target IgE at various concentrations caused a sharp decrease in the source-drain current, and a gradual saturation was observed at lower concentrations. The amount of the net source-drain current before and after IgE introduction on the aptamer-modified CNT-FETs increased as a function of IgE concentration. The detection limit for IgE was determined as 250 pM. We have also prepared CNT-FET biosensors using a monoclonal antibody against IgE (IgE-mAb). The electrical properties of the aptamer- and antibody-modified CNT-FETs were compared. The performance of aptamer-modified CNT-FETs provided better results than the ones obtained using IgE-mAb-modified CNT-FETs under similar conditions. Thus, we suggest that the aptamer-modified CNT-FETs are promising candidates for the development of label-free protein biosensors.

Label-free immunosensor for prostate-specific antigen based on single-walled carbon nanotube array-modified microelectrodes.

We have fabricated a label-free electrochemical immunosensor using microelectrode arrays modified with single-walled carbon nanotubes (SWNTs). Label-free detection of a cancer marker, total prostate-specific antigen (T-PSA), was carried out using differential pulse voltammetry (DPV). The current signals, derived from the oxidation of tyrosine (Tyr), and tryptophan (Trp) residues, increased with the interaction between T-PSA on T-PSA-mAb covalently immobilized on SWNTs. The selectivity of our biosensor was challenged using bovine serum albumin (BSA) as the target protein. The detection limit for T-PSA was determined as 0.25 ng/mL. Since the cut-off limit of T-PSA between prostate hyperplasia and cancer is 4 ng/mL, the performance of our label-free electrochemical immunosensor seems promising for further clinical applications.