Simultaneous Detection of Infectious Diseases Using Aptamer-Conjugated Gold Nanoparticles in the Lateral Flow Immunoassay-Based Signal Amplification Platform.

Various platforms for the accurate diagnosis of infectious diseases have been studied because of the emergence of coronavirus disease (COVID-19) in 2019. Recently, it has become difficult to distinguish viruses with similar symptoms due to the continuous mutation of viruses, and there is an increasing need for a diagnostic method to detect them simultaneously. Therefore, we developed a paper-based rapid antigen diagnostic test using DNA aptamers for the simultaneous detection of influenza A, influenza B, and COVID-19. Aptamers specific for each target viral antigen were selected and attached to AuNPs for application in a rapid antigen diagnosis kit using our company's heterogeneous sandwich-type aptamer screening method (H-SELEX). We confirmed that the three viruses could be detected on the same membrane without cross-reactivity based on the high stability, specificity, and binding affinity of the selected aptamers. Further, the limit of detection was 2.89 pg·mL-1 when applied to develop signal amplification technology; each virus antigen was detected successfully in diluted nasopharyngeal samples. We believe that the developed simultaneous diagnostic kit, based on such high accuracy, can distinguish various infectious diseases, thereby increasing the therapeutic effect and contributing to the clinical field.

Electrochemiluminescence-Incorporated Lateral Flow Immunosensors Using Ru(bpy)32+-Labeled Gold Nanoparticles for the Full-Range Detection of Physiological C-Reactive Protein Levels.

C-reactive protein (CRP) is used as a general biomarker for inflammation and infection. During stroke and myocardial infarction, CRP increases and is present in a broad concentration range of 1-500 µg/mL. Therefore, full-range CRP detection is crucial to identify patients who need close follow-up or intensive treatment after a heart attack. Here, we report the first attempt to develop an electrochemiluminescent lateral flow immunosensor (ECL-LFI) that allows full-range CRP detection. Ru(bpy)32+-labeled gold nanoparticles (AuNPs) are used as a CRP-targeting probe and a signal generator; they form sandwich immunocomplexes at the test line of the strip and generate strong ECL emission via a Ru(bpy)32+/tripropylamine system. The ECL-LFI shows high sensitivity in detecting CRP in spiked serum, with a limit of detection of 4.6 pg/mL within 15 min, and a broad detection range of 0.01-1000 ng/mL, which is 2 orders of magnitude broader than that of conventional colorimetric LFI. The clinical usability of the ECL-LFI was evaluated using 30 clinical serum samples (200 ng/mL to 5 mg/mL), which showed a good linear correlation (R2 = 0.9896), with a clinical chemistry analyzer. The results suggest that the ECL-LFI holds great potential for CRP detection in point-of-care diagnostics.

Ru(bpy)32+ -Loaded Mesoporous Silica Nanoparticles as Electrochemiluminescent Probes of a Lateral Flow Immunosensor for Highly Sensitive and Quantitative Detection of Troponin I.

The lateral flow immunosensor (LFI) is a widely used diagnostic tool for biomarker detection; however, its sensitivity is often insufficient for analyzing targets at low concentrations. Here, an electrochemiluminescent LFI (ECL-LFI) is developed for highly sensitive detection of troponin I (TnI) using Ru(bpy)32+ -loaded mesoporous silica nanoparticles (RMSNs). A large amount of Ru(bpy)32+ is successfully loaded into the mesoporous silica nanoparticles with excellent loading capacity and shows strong ECL signals in reaction to tripropylamine. Antibody-immobilized RMSNs are applied to detect TnI by fluorescence and ECL analysis after a sandwich immunoassay on the ECL-LFI strip. The ECL-LFI enables the highly sensitive detection of TnI-spiked human serum within 20 min at femtomolar levels (≈0.81 pg mL-1 ) and with a wide dynamic range (0.001-100 ng mL-1 ), significantly outperforming conventional fluorescence detection (>3 orders of magnitude). Furthermore, TnI concentrations in 35 clinical serum samples across a low range (0.01-48.31 ng mL-1 ) are successfully quantified with an excellent linear correlation (R2  = 0.9915) using a clinical immunoassay analyzer. These results demonstrate the efficacy of this system as a high-performance sensing strategy capable of capitalizing on future point-of-care testing markets for biomolecule detection.

Rapid PCR kit: lateral flow paper strip with Joule heater for SARS-CoV-2 detection.

Polymerase chain reaction (PCR)-based diagnostic kits for point-of-care (POC) testing are highly desirable to prevent the spread of infectious diseases. Here, we demonstrate a rapid PCR testing kit that involves integrating a lateral flow paper strip with a nichrome-based thin film heater. The use of a paper membrane as a PCR-solution container results in fast thermocycling without a cooler because the membrane can contain the solution with a high specific surface area where Joule heating is applied. After PCR, amplified products are simultaneously detected at the lateral flow paper strip with the naked eye. Severe acute respiratory syndrome ß-coronavirus RNA can be detected within 30 min after PCR solution injection. This work reveals that the paper membrane can act as not only a capillary flow channel but also as a promising platform for fast PCR and detection.

Plasmon color-preserved gold nanoparticle clusters for high sensitivity detection of SARS-CoV-2 based on lateral flow immunoassay.

Lateral flow immunoassays (LFI) have shown great promise for point-of-care (POC) sensing applications, however, its clinical translation is often hindered by insufficient sensitivity for early detection of diseases, including severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). This is mainly due to weak absorption signals of single gold nanoparticles (AuNPs). Here, we developed AuNP clusters that maintain the red color of isolated individual AuNPs, but increase the colorimetric readout to improve the detection sensitivity. The plasmon color-preserved (PLASCOP) AuNP clusters is simply made by mixing streptavidin-coated AuNP core with satellite AuNPs coated with biotinylated antibodies. The biotinylated antibody-streptavidin linker forms a gap size over 15 nm to avoid plasmon coupling between AuNPs, thus maintaining the plasmonic color while increasing the overall light absorption. LFI sensing using PLASCOP AuNP clusters composed of 40 nm AuNPs showed a high detection sensitivity for SARS-CoV-2 nucleocapsid proteins with a limit of detection (LOD) of 0.038 ng mL-1, which was 23.8- and 5.9-times lower value than that of single 15 nm and 40 nm AuNP conjugates, respectively. The PLASCOP AuNP clusters-based LFI sensing also shows good specificity for SARS-CoV-2 nucleocapsid proteins from other influenza and coronaviruses. In a clinical feasibility test, we demonstrated that SARS-CoV-2 particles spiked in human saliva could be detected with an LOD of 54 TCID50 mL-1. The developed PLASCOP AuNP clusters are promising colorimetric sensing reporters that present improved sensitivity in LFI sensing for broad POC sensing applications beyond SARS-CoV-2 detection.

Physisorption of Affinity Ligands Facilitates Extracellular Vesicle Detection with Low Non-Specific Binding to Plasmonic Gold Substrates.

Plasmonic biosensors are increasingly being used for the analysis of extracellular vesicles (EVs) originating from disease areas. However, the high non-specific binding of EVs to a gold-sensing surface has been a critical problem and hindered the true translational potential. Here, we report that direct antibody immobilization on the plasmonic gold surface via physisorption shows excellent capture of cancer-derived EVs with ultralow non-specific binding even at very high concentrations. Contrary to commonly used methods that involve thiol-based linker attachment and an EDC/sulfo-NHS reaction, we show a higher specific capture rate and >50-fold lower non-specific on citrate-capped plain and nanopatterned gold surfaces. The method provides a simple, fast, and reproducible means to functionalize plasmonic gold surfaces with antibodies for robust EV biosensing.

Wafer-Scale LSPR Substrate: Oblique Deposition of Gold on a Patterned Sapphire Substrate.

Label-free detection of biomolecules using localized surface plasmon resonance (LSPR) substrates is a highly attractive method for point-of-care (POC) testing. One of the remaining challenges to developing LSPR-based POC devices is to fabricate the LSPR substrates with large-scale, reproducible, and high-throughput. Herein, a fabrication strategy for wafer-scale LSPR substrates is demonstrated using reproducible, high-throughput techniques, such as nanoimprint lithography, wet-etching, and thin film deposition. A transparent sapphire wafer, on which SiO2-nanodot hard masks were formed via nanoimprint lithography, was anisotropically etched by a mixed solution of H2SO4 and H3PO4, resulting in a patterned sapphire substrate (PSS). An LSPR substrate was finally fabricated by oblique deposition of Au onto the PSS, which was then applied to label-free detection of the binding events of biomolecules. To the best of our knowledge, this paper is the first report on the application of the PSS used as an LSPR template by obliquely depositing a metal.

Advanced trap lateral flow immunoassay sensor for the detection of cortisol in human bodily fluids.

Paper-based biosensors based on lateral flow immunoassay (LFI) are promising candidates for POC diagnosis because of their ease of use and rapid target detection. However, the low sensitivity of LFI limits its application, and signal amplification has been used in numerous studies to increase its sensitivity. We developed an advanced trap LFI (α-trapLFI), a simple-to-use sensor, with an additional step for signal amplification. Here, signal amplification is automatically implemented following delayed release of enhancement solution induced by water-soluble polyvinyl alcohol tape. As the polyvinyl alcohol tape is exposed to water, its polymer structure is perturbed (within 5 min), allowing ions to pass through. This new sensor was designed to have a short time delay between the flow of solutions used for the immunoassay and signal amplification. The α-trapLFI was subsequently used to detect cortisol with high sensitivity (9.1 pg∙mL-1) over a broad detection range (0.01-1000 ng∙mL-1) in bodily fluids. Furthermore, an excellent correlation was obtained by analyzing 20 human real saliva samples using this sensor and a conventional ELISA (R2 = 0.90). The new sensor will be helpful in detecting various small molecules for simple, rapid, and portable POC diagnosis of stress disorders.

Reactant/polymer hybrid films on p-n junction photodetectors for self-powered, non-invasive glucose biosensors.

The portability of electronic-based biosensors is limited because of the use of batteries and/or solutions containing reactants such as enzymes for assay, which limits the utility of such biosensors in point-of-care (POC) testing. In this study, we report on the development of a self-powered biosensor composed of only portable components: a reactant-containing poly (ethylene glycol) (PEG) film for the colorimetric assay, and a self-powered n-InGaZnO/p-Si photodetector. The PEG film containing enzymes and color-developing agents was formed on a glass slide by spin coating. The self-powered biosensor was fabricated by placing the hybrid film on the p-n junction photodetector, and applied in non-invasive glucose detection (salivary glucose). Injection of the target-containing solution dissolved the PEG that led to the release of enzymes and color-developing agents, resulting in a colorimetric assay. The colorimetric assay could attenuate the light reaching the photodetector, thus facilitating target concentration verification by measuring the photocurrent. Our self-powered biosensor has two main advantages: (i) all components of the biosensor are portable and (ii) dilution of target concentration is avoided as the reagents are in the PEG film. Therefore, the self-powered biosensor, without solution-phase components, could be highly beneficial for creating portable, sensitive biosensors for POC testing.

Open-Shell and Closed-Shell Quinoid-Aromatic Conjugated Polymers: Unusual Spin Magnetic and High Charge Transport Properties.

While quinoidal moieties are considered as emerging platforms showing efficient charge transport and interesting open-shell diradical characteristics, whether these properties could be changed by extension to the conjugated polymer structure remains as a fundamental question. Here, we developed and characterized two conjugated polymers incorporating quinoids with different lengths, which have a stable close- and open-shell diradical character, respectively, namely, poly(quinoidal thiophene-thienylene vinylene) (PQuT-TV) and poly(quinoidal bithiophene-thienylene vinylene) (PQuBT-TV). A longer length of a quinoidal core led to enhanced diradical characteristics. Therefore, the longer core length of QuBT was favorable for the formation of an open-shell diradical structure in its monomer and in the quinoidal polymer. PQuBT-TV exhibited high spin characteristics observed by the strong ESR signal, a low band gap, and improved electrochemical stability. On the other hand, as QuT maintained a closed-shell quinoid structure, PQuT-TV exhibited high backbone coplanarity and strong intermolecular interaction, which was beneficial for charge transport and led to high hole mobility (up to 2.40 cm2 V-1 s-1) in organic field-effect transistors. This work successfully demonstrated how the control of the closed/open-shell character of quinoidal building blocks changes charge transport and spin properties of quinoidal conjugated polymers via quinoid-aromatic interconversion.

Gold nanocap-supported upconversion nanoparticles for fabrication of a solid-phase aptasensor to detect ochratoxin A.

Solid-phase, single-step biosensors are crucial for the development of portable, reusable, and convenient biosensors, otherwise known as point-of-care (POC) testing. Although high-performance single-step biosensors based on the principle of Förster resonance energy transfer (FRET) and using upconversion nanoparticles (UCNPs) functionalized with aptamers have been suggested as easy-to-use platforms, they lack portability and reusability when used for solution-phase biosensing. In this study, we describe a solid-phase, single-step aptasensor that showed higher performance than those of solution-phase aptasensors, as well as promising reusability. The solid-phase, single-step aptasensor was developed based on Au nanocap-supported UCNPs (Au/UCNPs), which were partially embedded in a solid substrate (e.g. polydimethylsiloxane, PDMS). The Au nanocaps allowed the UCNPs to emit upconverted light only from the restricted areas of the UCNPs, i.e., where they were not covered by the nanocaps and PDMS. Functionalization of an aptamer labeled with a quencher on the restricted area enabled the effective quenching of upconverted light from Au/UCNP via FRET after target (ochratoxin A, OTA) detection. The solid-phase, single-step aptasensor showed a linear range of 0.1-1000 ng mL-1 and limit of detection of 0.022 ng mL-1 within 30 min toward OTA. Furthermore, reusability of the solid-phase aptasensor was evaluated for three cycles of detection and regeneration, establishing its apparent reusability via heat treatment. Hence, such solid-phase, single-step aptasensors pave the path to the development of a portable and reusable biosensor platform for POC testing.

Advanced Colorimetric Paper Sensors Using Color Focusing Effect Based on Asymmetric Flow of Fluid.

Although paper-based colorimetric sensors utilizing enzymatic reactions are well suited for real-field diagnosis, their widespread use is hindered by signal blurring at the detection spot due to the action of capillary forces on the liquid and the corresponding membrane. In this study, we eliminated signal losses commonly observed during enzyme-mediated colorimetric sensing and achieved pattern-free quantitative analysis of glucose and uric acid by mixing enzymes and color-forming reagents with chitosan oligosaccharide lactate (COL), which resulted in perfectly focused colorimetric signals at the detection spot, using asymmetric flow induced by changing the flow rate of the COL-treated paper. The targets were calibrated with 0-500 mg/dL of glucose and 0-200 mg/dL of uric acid, and the limit of detection was calculated to be 0.6 and 0.03 mg/dL, respectively. In human urine, the correlation has a high response between the measured and spiked concentrations, and the stability of the enzyme mixture including COL increased by 41% for glucose oxidase mixture and 29% for uricase mixture, compared to the corresponding mixtures without COL. Thus, the color focusing and pattern-free sensor, which have the advantages of easy fabrication, easy handling, and high stability, should be applied to real-field diagnosis.

Plasmon enhanced up-conversion nanoparticles in perovskite solar cells for effective utilization of near infrared light.

As an alternative to silicon-based solar cells, organic-inorganic hybrid perovskite solar cells (PSCs) have attracted much attention and achieved a comparable power conversion efficiency (PCE) to silicon-based ones, although the perovskite materials can absorb only visible light. Hence, the challenge remains to enhance the PCE utilizing near infrared (NIR) light in the solar light spectrum. One of the easiest ways to utilize the NIR is to incorporate NIR active materials in PSCs such as up-conversion nanoparticles (UCNPs); however, such a stratergy is not simple to adopt in PSCs due to the inherent vurnerability of perovskite materials towards moisture. In this work, we present NIR-utilizing PSCs by locating UCNPs within the PSC structure by a simple dry transfer method. A maximum PCE of 15.56% was obtained in the case of PSC having the UCNPs located between the hole transport layer (HTL) and gold (Au) top electrode, which is an 8.4% enhancement compared to the cell without the UCNPs. This enhancement came from the combined effects of NIR light utilization and the surface plasmon resonance (SPR) phenomenon originating from the Au top electrode, which was interfacing the UCNPs.

Self-Powered Biosensors Using Various Light Sources in Daily Life Environments: Integration of p-n Heterojunction Photodetectors and Colorimetric Reactions for Biomolecule Detection.

Electronic biosensors operating without power supply are high in demand owing to increasing interest in point-of-care (POC) coupled with portable and wearable electronic devices for smart healthcare services. Although self-powered electronic sensors have emerged with the promise of resolving the energy supply problems, achieving sufficient sensitivity to targets in real samples is highly challenging because of the matrix effect caused by electroactive species. In this study, we developed a self-powered biosensor platform by combining n-indium gallium zinc oxide (IGZO)/p-Si heterojunction photodetectors and physically separated colorimetric reactions. The self-powered biosensors were applied to glucose detection in real human samples using light sources from daily life environments such as fluorescent light and sunlight. The sensors showed high sensitivity and stability from 0.01 to 10 mg mL-1 of glucose in human saliva and urine without matrix effect from the electroactive species in real samples. In addition, a small change in glucose concentration in human serum was distinguishable with a resolution of 0.01 mg mL-1. Notably, these results were obtained using well-developed and widely used materials like Si and IGZO with simple deposition techniques. Moreover, this self-powered biosensing platform can be universally applied for the detection of all biomolecules being detected by colorimetric assays. To the best of our knowledge, this is the first report on such self-powered biosensors, which could be a promising candidate for future POC biosensors integrated with portable and wearable electronic devices.

Low-temperature-grown continuous graphene films from benzene by chemical vapor deposition at ambient pressure.

There is significant interest in synthesizing large-area graphene films at low temperatures by chemical vapor deposition (CVD) for nanoelectronic and flexible device applications. However, to date, low-temperature CVD methods have suffered from lower surface coverage because micro-sized graphene flakes are produced. Here, we demonstrate a modified CVD technique for the production of large-area, continuous monolayer graphene films from benzene on Cu at 100-300 °C at ambient pressure. In this method, we extended the graphene growth step in the absence of residual oxidizing species by introducing pumping and purging cycles prior to growth. This led to continuous monolayer graphene films with full surface coverage and excellent quality, which were comparable to those achieved with high-temperature CVD; for example, the surface coverage, transmittance, and carrier mobilities of the graphene grown at 300 °C were 100%, 97.6%, and 1,900-2,500 cm(2) V(-1) s(-1), respectively. In addition, the growth temperature was substantially reduced to as low as 100 °C, which is the lowest temperature reported to date for pristine graphene produced by CVD. Our modified CVD method is expected to allow the direct growth of graphene in device manufacturing processes for practical applications while keeping underlying devices intact.

Palladium Nanoribbon Array for Fast Hydrogen Gas Sensing with Ultrahigh Sensitivity.

A lithographically aligned palladium nano-ribbon (Pd-NRB) array with gaps of less than 40 nm is fabricated on a poly(ethylene terephthalate) substrate using the direct metal transfer method. The 200 µm Pd-NRB hydrogen gas sensor exhibits an unprecedented sensitivity of 10(9) % after bending treatment, along with fast sensing behavior (80% response time of 3.6 s and 80% recovery time of 8.7 s) at room temperature.