Direct electron transfer (DET) between an electrode and redox labels is feasible in electrochemical biosensors using small aptamer-aptamer sandwiches; however, its application is limited in biosensors that rely on larger antibody-antibody sandwiches. The development of sandwich-type biosensors utilizing DET is challenged by the scarcity of aptamer-aptamer sandwich pairs with high affinity in complex biological samples. Here, we introduce an electrochemical biosensor using an antibody-aptamer hybrid sandwich for detecting thrombin in human serum.
View Article and Find Full Text PDFTo minimize background interference in electrochemical enzymatic biosensors employing electron mediators, it is essential for the electrochemical oxidation of electroactive interfering species (ISs), such as ascorbic acid (AA), to proceed slowly, and for the redox reactions between electron mediators and ISs to occur at a low rate. In this study, we introduce a novel combination of a working electrode and an electron mediator that effectively mitigates interference effects. Compared to commonly used electrodes such as Au, glassy carbon, and indium tin oxide (ITO), boron-doped diamond (BDD) electrodes demonstrate significantly lower anodic current (i.
View Article and Find Full Text PDFThe selective electrochemical synthesis of 1H-indazoles and their N-oxides and the subsequent C-H functionalization of the 1H-indazole N-oxides are described. The electrochemical outcomes were determined by the nature of the cathode material. When a reticulated vitreous carbon cathode was used, a wide range of 1H-indazole N-oxides were selectively synthesized, and the electrosynthesis products were deoxygenated to N-heteroaromatics, owing to cathodic cleavage of the N-O bond via paired electrolysis, when a Zn cathode was used.
View Article and Find Full Text PDFAn electron mediator with rapid dissolvability and high solubility in aqueous electrolyte solutions is essential for point-of-care testing based on mediated electrochemical detection. However, most ferrocenyl (Fc) compounds have slow dissolvability and poor solubility owing to high hydrophobicity of the Fc backbone. Moreover, many Fc compounds have poor stability and nonoptimal formal potential ().
View Article and Find Full Text PDFDirect electron transfer (DET) between a redox label and an electrode has been used for sensitive and selective sandwich-type detection without a wash step. However, applying DET is still highly challenging in protein detection, and a single redox label per probe is insufficient to obtain a high electrochemical signal. Here, we report a wash-free, sandwich-type detection of thrombin using DET and catalytic signal amplification of multiple redox labels.
View Article and Find Full Text PDFThe commonly required properties of diffusive electron mediators for point-of-care testing are rapid dissolubility, high stability, and moderate formal potential in aqueous solutions. Inspired by nature, various quinone-containing electron mediators have been developed; however, satisfying all these requirements remains a challenge. Herein, a strategic design toward quinones incorporating sulfonated thioether and nitrogen-containing heteroarene moieties as solubilizing, stabilizing, and formal potential-modulating groups is reported.
View Article and Find Full Text PDFBiosens Bioelectron
May 2021
Recombinase polymerase amplification (RPA) is considered one of the best amplification methods for realizing a miniaturized diagnostic instrument; however, it is notably challenging to obtain low detection limits in solid-phase RPA. To overcome these difficulties, we combined solid-phase RPA with electrochemical detection and used a new concentration combination of three primers (surface-bound forward primer, solution reverse primer, and an extremely low concentration of solution forward primer). When solid-phase RPA was performed on an indium tin oxide (ITO) electrode modified with a surface-bound forward primer in a solution containing a biotin-terminated solution reverse primer, an extremely low concentration of a solution forward primer, and a template DNA or genomic DNA for a target gene of hepatitis B virus (HBV), amplification occurred mainly in solution until all the solution forward primers were consumed.
View Article and Find Full Text PDFThe duplex detection of both total and active enzyme concentrations without interferences at a single working electrode is challenging, especially when two different assays are combined. It is also challenging to obtain two different redox-cycling reactions without interference. Here, we present a simple but sensitive combined assay that is based on two redox-cycling reactions using two incubation periods and applied potentials at a single electrode.
View Article and Find Full Text PDFMetal nanoparticle surfaces are used for peroxidase- and oxidase-like nanozymes but not for esterase-like nanozymes. It is challenging to obtain rapid catalytic hydrolysis on a metal surface and even more so without a catalytically labile substrate. Here, we report that metal nanoparticle surfaces rapidly catalyze non-redox ester hydrolysis in the presence of redox H N-BH (AB).
View Article and Find Full Text PDFPropagating cascade reactions based on two proteases are promising for obtaining high signal amplification. However, in many cases, biosensors that use cascade reactions do not have low detection limits because of the inherent slowness of proteolytic reactions. Here, we report a sensitive electrochemical immunosensor using a high-signal-amplification method that combines a propagating cascade reaction and a redox cycling reaction.
View Article and Find Full Text PDF