Surface Modification of Screen-Printed Carbon Electrode through Oxygen Plasma to Enhance Biosensor Sensitivity.

The screen-printed carbon electrode (SPCE) is a useful technology that has been widely used in the practical application of biosensors oriented to point-of-care testing (POCT) due to its characteristics of cost-effectiveness, disposability, miniaturization, wide potential window, and simple electrode design. Compared with gold or platinum electrodes, surface modification is difficult because the carbon surface is chemically or physically stable. Oxygen plasma (O2) can easily produce carboxyl groups on the carbon surface, which act as scaffolds for covalent bonds. However, the effect of O2-plasma treatment on electrode performance remains to be investigated from an electrochemical perspective, and sensor performance can be improved by clarifying the surface conditions of plasma-treated biosensors. In this research, we compared antibody modification by plasma treatment and physical adsorption, using our novel immunosensor based on gold nanoparticles (AuNPs). Consequently, the O2-plasma treatment produced carboxyl groups on the electrode surface that changed the electrochemical properties owing to electrostatic interactions. In this study, we compared the following four cases of SPCE modification: O2-plasma-treated electrode/covalent-bonded antibody (a); O2-plasma-treated electrode/physical adsorbed antibody (b); bare electrode/covalent-bonded antibody (c); and bare electrode/physical absorbed antibody (d). The limits of detection (LOD) were 0.50 ng/mL (a), 9.7 ng/mL (b), 0.54 ng/mL (c), and 1.2 ng/mL (d). The slopes of the linear response range were 0.039, 0.029, 0.014, and 0.022. The LOD of (a) was 2.4 times higher than the conventional condition (d), The slope of (a) showed higher sensitivity than other cases (b~d). This is because the plasma treatment generated many carboxyl groups and increased the number of antibody adsorption sites. In summary, the O2-plasma treatment was found to modify the electrode surface conditions and improve the amount of antibody modifications. In the future, O2-plasma treatment could be used as a simple method for modifying various molecular recognition elements on printed carbon electrodes.

Wireless electrochemiluminescent biosensors: Powering innovation with smartphone technology.

Energy supply and sensor response acquisition can be performed wirelessly, enabling biosensors as Internet of Thing (IoT) tools by linking wireless power supply and electrochemical sensors. Here, we used the electromagnetic induction method to clarify the conditions under which electrochemiluminescence is induced by a simple potential modulation circuit without an integrated circuit on the electrode chip that receives the power. Initially, the potential waveform obtained in a circuit with inductance and capacitance components that resonate with the transmission frequency and a diode for rectification was investigated to clarify the conditions inducing an electrochemiluminescence reaction at the printed electrode. A high-sensitivity complementary metal-oxide semiconductor camera built into the smartphone wirelessly detected the luminescence generated on the electrode chip. The images were quantitatively evaluated using open-source image analysis software which determine the sensitivity of detecting hydrogen peroxide. Glucose oxidase (GOD) encapsulated in a matrix of chitosan polymers and photocrosslinkable polymers was immobilized on a mass-producible and inexpensive printed electrode to maintain high activity. The immobilized membrane suppressed luminescence when immobilized on the working electrode; therefore, the enzyme was immobilized on the counter electrode for glucose measurement over a wide concentration. Thus, luminol electrochemiluminescence was induced on the electrode chip by wireless power supply from a smartphone. Human serum and artificial sweat samples were tested and indicated possibility for actual applications. In this way a fully wireless biosensor was developed with potential as an IoT biosensor.

Point-of-Care Diagnostic Biosensors to Monitor Anti-SARS-CoV-2 Neutralizing IgG/sIgA Antibodies and Antioxidant Activity in Saliva.

Monitoring biomarkers is a great way to assess daily physical condition, and using saliva instead of blood samples is more advantageous as the process is simple and allows individuals to test themselves. In the present study, we analyzed the titers of neutralizing antibodies, IgG and secretory IgA (sIgA), in response to the SARS-CoV-2 vaccine, in saliva. A total of 19 saliva and serum samples were collected over a 10-month period 3 weeks after the first vaccine, 8 months after the second vaccine, and 1 month after the third vaccine. The ranges of antibody concentrations post-vaccination were: serum IgG: 81-15,000 U/mL, salivary IgG: 3.4-330 U/mL, and salivary IgA: 58-870 ng/mL. A sharp increase in salivary IgG levels was observed after the second vaccination. sIgA levels also showed an increasing trend. A correlation with trends in serum IgG levels was observed, indicating the possibility of using saliva to routinely assess vaccine efficacy. The electrochemical immunosensor assay developed in this study based on the gold-linked electrochemical immunoassay, and the antioxidant activity measurement based on luminol electrochemiluminescence (ECL), can be performed using portable devices, which would prove useful for individual-based diagnosis using saliva samples.

Fully Transparent, Ultrathin Flexible Organic Electrochemical Transistors with Additive Integration for Bioelectronic Applications.

Optical transparency is highly desirable in bioelectronic sensors because it enables multimodal optical assessment during electronic sensing. Ultrathin (<5 µm) organic electrochemical transistors (OECTs) can be potentially used as a highly efficient bioelectronic transducer because they demonstrate high transconductance during low-voltage operation and close conformability to biological tissues. However, the fabrication of fully transparent ultrathin OECTs remains a challenge owing to the harsh etching processes of nanomaterials. In this study, fully transparent, ultrathin, and flexible OECTs are developed using additive integration processes of selective-wetting deposition and thermally bonded lamination. These processes are compatible with Ag nanowire electrodes and conducting polymer channels and realize unprecedented flexible OECTs with high visible transmittance (>90%) and high transconductance (≈1 mS) in low-voltage operations (<0.6 V). Further, electroencephalogram acquisition and nitrate ion sensing are demonstrated in addition to the compatibility of simultaneous assessments of optical blood flowmetry when the transparent OECTs are worn, owing to the transparency. These feasibility demonstrations show promise in contributing to human stress monitoring in bioelectronics.

Towards On-site Determination of Secretory IgA in Artificial Saliva with Gold-Linked Electrochemical Immunoassay (GLEIA) Using Portable Potentiostat and Disposable Printed Electrode.

Mental stress is closely connected with our physical and mental wellness. Therefore, stress measurement can contribute to assess our lifestyle and increase our quality of life. In this paper, we detect the secretory immunoglobulin A (sIgA), which is the candidate of salivary stress markers, with original electrochemical immunoassay: gold-linked electrochemical immunoassay (GLEIA). This biosensor is based on a sandwich-type immunosensor and adopts the electrochemical method to detect the reduction peak from Au nanoparticles linked to the secondary antibody. GLEIA is convenient and cost-effective that only requires a low sample volume (10 µL). In addition, the GLEIA show high sensitivity and selectivity. We obtained the linear response to relate the concentration of sIgA (10-300 ng/mL) in D-PBS buffer with the artificial saliva which includes salivary inorganic salt and typically glycoprotein (mucin). Furthermore, we obtained acceptable selectivity in the various solution with salivary proteins such as α-amylase, human serum albumin, immunoglobulin G (IgG), lysozyme, and mucin. In the future, we try to detect the sIgA in real saliva for on-site stress measurement using GLEIA and to integrate the various immunosensors for stress markers in saliva.

Investigation of Polyurethane Matrix Membranes for Salivary Nitrate ISFETs to Prevent the Drift.

We have investigated human-stress monitoring by making use of salivary nitrate, which can be a candidate for stress markers, with ion-selective field-effect transistors (ISFETs). ISFETs are suitable for on-site single-drop analysis of salivary nitrate within 10 s. However, when ISFETs are used for salivary nitrate, ISFETs have a problem that is called the initial drift. The initial drift makes accurate nitrate monitoring difficult. Thus, the purpose of this study is to prevent the initial drift and to search for a new, simple polymer to possess a better performance of sensor responses than conventional matrix membranes, such as PVC. In this research, we investigated ISFETs using specific matrix membranes, for example KP-13, Pellethane®--, and P7281-PU. The initial drift was evaluated from the fluctuations of the response values generated by the ISFETs when immersed in saliva or aqueous solution. As a result, P7281-PU showed a prevention effect on the initial drift, both in the whole saliva and in various solutions. Furthermore, the cause of drift may be H+ diffusion, and the drift prevention effect of P7281-PU may be affected by urethane bond capturing H+ in the ion-selective membrane. This result suggests that a continuous nitrate monitoring is feasible and may be applied to wearable sensors.