Multiwalled Carbon Nanotube-Templated Nickel Porphyrin Covalent Organic Framework for Pencil-Drawn Noninvasive Respiration Sensors.

Paper-integrated configuration with miniaturized functionality represents one of the future main green electronics. In this study, a paper-based respiration sensor was prepared using a multiwalled carbon nanotube-templated nickel porphyrin covalent organic framework (MWCNTs@COFNiP-Ph) as an electrical identification component and pencil-drawn graphite electric circuits as interdigitated electrodes (IDEs). The MWCNTs@COFNiP-Ph not only inherited the high gas sensing performance of porphyrin and the aperture induction effect of COFs but also overcame the shielding effect between phases through the MWCNT template. Furthermore, it possessed highly exposed M-N4 metallic active sites and unique periodic porosity, thereby effectively addressing the key technical issue of room-temperature sensing for the respiration sensor. Meanwhile, the introduction of a pencil-drawing approach on common printing papers facilitates the inexpensive and simple manufacturing of the as-fabricated graphite IDE. Based on the above advantages, the MWCNTs@COFNiP-Ph respiration sensor had the characteristics of wide detection range (1-500 ppm), low detection limit (30 ppb), acceptable flexibility for toluene, and rapid response/recovery time (32 s/116 s). These advancements facilitated the integration of the respiration sensor into surgical masks and clothes with maximum functionality at a minimized size and weight. Moreover, the primary internal mechanism of COFNiP-Ph for this efficient toluene detection was investigated through in situ FTIR spectra, thereby directly elucidating that the chemisorption interaction of oxygen modulated the depletion layers, resulting in alterations in sensor resistance upon exposure to the target gas. The encouraging results revealed the feasibility of employing a paper-sensing system as a wearable platform in green electronics.

Double-enzyme active MnO2@BSA mediated lab-on-paper dual-modality aptasensor for di(2-ethylhexyl)phthalate.

Di(2-ethylhexyl)phthalate (DEHP), as an environmental endocrine disruptor, has adverse effects on eco-environments and health. Thus, it is crucial to highly sensitive on-site detect DEHP. Herein, a double-enzyme active MnO2@BSA mediated dual-modality photoelectrochemical (PEC)/colorimetric aptasensing platform with the cascaded sensitization structures of ZnIn2S4 and TiO2 as signal generators was engineered for rapid and ultrasensitive detection of DEHP using an all-in-one lab-on-paper analytical device. Benefitting from cascaded sensitization effect, the ZnIn2S4/TiO2 photosensitive structures-assembled polypyrrole paper electrode gave an enhanced photocurrent signal. The MnO2@BSA nanoparticles (NPs) with peroxidase-mimic and oxidase-mimic double-enzymatic activity induced multiple signal quenching effects and catalyzed color development. Specifically, the MnO2@BSA NPs acted as peroxidase mimetics to generate catalytic precipitates, which not only obstructed interfacial electron transfer but also served as electron acceptors to accept photogenerated electrons. Besides, the steric hindrance effect from MnO2@BSA NPs-loaded branchy polymeric DNA duplex structures further decreased photocurrent signal. The target recycling reaction caused the detachment of MnO2@BSA NPs to increase PEC signal, realizing the ultrasensitive detection of DEHP with a low detection limit of 27 fM. Ingeniously, the freed MnO2@BSA NPs flowed to colorimetric zone with the aid of fluid channels and acted as oxidase mimetics to induce color intensity enhancement, resulting in the rapid visual detection of DEHP. This work provided a prospective paradigm to develop field-based paper analytical tool for DEHP detection in aqueous environment.

Rolling circle amplification assisted CRISPR/Cas12a dual-cleavage photoelectrochemical biosensor for highly sensitive detection of miRNA-21.

BACKGROUND: MicroRNA-21 has been determined to be the only microRNA overexpressed in 11 types of solid tumors, making it an excellent candidate as a biomarker for disease diagnosis and therapy. Photoelectrochemical (PEC) biosensors have been widely used for quantification of microRNA-21. However, most PEC biosensing processes still suffer from some problems, such as the difficulty of avoiding the influence of interferents in complex matrices and the false-positive signals. There is a pressing need for establishing a sensitive and stable PEC method to detect microRNA-21. RESULTS: Herein, a nicking endonuclease-mediated rolling circle amplification (RCA)-assisted CRISPR/Cas12a PEC biosensor was fabricated for ultrasensitive detection of microRNA-21. The p-p type heterojunction PbS QDs/Co3O4 polyhedra were prepared as the quencher, thus the initial PEC signal attained the "off" state. Furthermore, the target was specifically identified and amplified by the RCA process. Then, its product single-stranded DNA S1 activated the cis- and trans-cleavage abilities of CRISPR/Cas12a, leading to almost all of the PbS QDs/Co3O4 polyhedra to leave the electrode surface, the p-n semiconductor quenching effect to be disrupted, and the signal achieving the "super-on" state. This pattern of PEC signal changed from "off" to "on" eliminated the interference of false-positive signals. The proposed PEC biosensor presented a satisfactory linear relationship ranging from 1 fM to 10 nM with a detection limit of 0.76 fM (3 Sb/N). SIGNIFICANCE AND NOVELTY: With innovatively synthesized PbS QDs/Co3O4 polyhedra as the effective quencher for PEC signal, the CRISPR/Cas12a dual-cleavage PEC biosensor possessed excellent selectivity, stability and repeatability. Furthermore, the detection of various miRNAs can be realized by changing the relevant base sequences in the constructed PEC biosensor. It also provides a powerful strategy for early clinical diagnosis and biomedical research.

Peptide cleavage-mediated photoelectrochemical signal on-off via CuS electronic extinguisher for PSA detection.

In this work, a peptide-based photoelectrochemical (PEC) biosensor was constructed based on CdTe/TiO2 sensitized structure as electrode and CuS nanocrystals as signal amplifier for the ultrasensitive detection of protein. After peptide was fixed to the CdTe/TiO2 electrode surface, the double-helix DNA (dsDNA) was immobilized at the end of the peptide and used as a carrier to immobilize the doxorubicin-copper sulfide nanocrystals (Dox-CuS) conjugates. As a proof of concept, prostate specific antigen (PSA) has been chosen as the model. In absence of PSA, CuS nanocrystals could consume electron donors and exciting light energy. Additionally, the steric hindrance effect of biomacromolecules hindered the movement of electron donors to the photoelectrode. Eventually, the photoelectric response signal was reduced, and the biosensor was in a "signal off" state. When PSA existed, the PSA specifically cleaved the peptide, and DNA/Dox-CuS probes were released from the electrode surface, resulting in a "signal on" state. The PEC biosensor revealed good specificity, stability, and reproducibility, and it exhibited excellent application in PSA analysis with a linear range from 0.005 to 20 ng mL-1 and a low detection limit of 0.0015 ng mL-1. This PEC biosensor may have potential applications in bioanalysis, disease diagnostics, and clinical biomedicine.

Donor/Acceptor-Induced Ratiometric Photoelectrochemical Paper Analytical Device with a Hollow Double-Hydrophilic-Walls Channel for microRNA Quantification.

Integrating ratiometric photoelectrochemical (PEC) techniques with paper microfluidics to construct a ratiometric PEC paper analytical device for practical application is often restricted by the grave dependence of ratiometric assay on photoactive materials and low mass-transfer rates of the paper channel. Herein, a universal donor/acceptor-induced ratiometric PEC paper analytical device with a hollow double-hydrophilic-walls channel (HDHC) was fabricated for high-performance microRNA-141 (miRNA-141) quantification. Concretely, a photoanode and photocathode were integrated on the paper-based sensing platform in which the photocathode served as a biosensing site for the pursuit of higher selectivity. For formulation of a cascading signal amplification strategy, a unique duplex-specific nuclease-induced target recycling reaction was engineered for the output of a double amount of all useful DNA linkers instead of conventional output of only one available DNA product, which could guarantee the output of abundant DNA linkers with the initiation of a cascade of hybridization chain reaction on both the trunk and branch in the presence of miRNA-141. Then the formed dendriform polymeric DNA duplex structures were further decorated with glucose oxidase (GOx)-mimicking gold nanoparticles by the electrostatic interaction to form a branchy gold tree (BGT). Profiting from the perfect GOx-mimicking activity of BGT and high mass-transfer rates of HDHC, the cathodic photocurrent from Ag2S/Cu2O hybrid structure was in a "signal off" state while the anodic photocurrent from graphene quantum dots (GQDs) and Ag2Se QDs cosensitized ZnO nanosheets was in a "signal on" state because BGT-catalyzed glucose oxidation reaction evoked the consumption of dissolved O2 as an electron acceptor and the generation of H2O2 as an electron donor. With calculation of the ratio of two photocurrent intensities, the quantitative detection of miRNA-141 was achieved with high sensitivity, accuracy, and reliability.

Triggerable H2O2-Cleavable Switch of Paper-Based Biochips Endows Precision of Chemometer/Ratiometric Electrochemical Quantification of Analyte in High-Efficiency Point-of-Care Testing.

In this work, a triggerable H2O2-cleavable fluid switch mediated paper-based biochip, being amenable to multiplexing and quantitative analysis with the dual-response output of visual screening and ratiometric electrochemistry, was developed for sensitive detection of target on-site. By properly implanting hydrophobic Ag-H2O2 responsive material in specific zone to form a programmable fluid switch, the biochip could achieve different modes of blocking/connecting switching automatically. In order to improve the test performance, a ratiometric electrochemical signal readout was designed, which was enhanced by a secondary in situ growth method fabricating trepang-shaped Au modified paper working electrode. In virtue of hybridization chain reaction, classic competitive recognition interactions of aptamer and target, and ratiometric internally calibrated mechanisms, ultrasensitive detection of the target was realized. To acquire a more quantitative and straightforward naked eye visual screening, the hydrophobic Ag switch was triggered by stimulating instructions from H2O2, thus reconnecting the electrochemical and ratiometric units automatically and resulting in a "signal on" visual fluidic flow on the chemometer characterized by the accurate distance of color development as a detection motif. With MCF-7 and K562 cells as models, wider linear detection ranges from 150 to 1.0 × 107 and 220 to 7.0 × 106 cells mL-1 for MCF-7 and K562 cells, respectively, were achieved. Meanwhile, thanks to the paper fluid chemometer, an acceptable screening detection limit of 103 cells mL-1 was obtained in the quantitative colorimetric assays. The proposed paper-based biochips opened up new horizons for designing of integratable, easy-to-use, and precise point-of-care testing devices.

Mimic peroxidase-transfer enhancement of photoelectrochemical aptasensing via CuO nanoflowers functionalized lab-on-paper device with a controllable fluid separator.

Inspired by the design of folding greeting cards and tissue drawing covers, a photoelectrochemical (PEC) lab-on-paper device with a controllable fluid separator, producing both reaction zone and detection zone, was explored for ultrasensitive detection of adenosine 5'-triphosphate (ATP) via mimic peroxidase-transfer enhancement of photocurrent response. To realize it, the DNA1, aptamer, and DNA2 as well as the mimic peroxidase of G-quadruplex/hemin modified Au nanocubes were linked on the graphene oxide-functionalized reaction zone via the DNA hybridization. Meanwhile, three-dimensional CuO nanoflowers (CuO NFs) as a photoactive material with outstanding electron transfer ability and absorption of light were grown in situ on the detection zone, providing a PEC active interface. Besides, an innovative fluid separator was elaborately designed by assembling a strip of paper with a hydrophilic channel, providing an effective way to bridge the gap between the two zones with a controllable drawing way, which could successfully avoid the signal interference caused by modifying biomolecules layer by layer on photosensitive materials. In the presence of ATP, the G-quadruplex/hemin modified in the reaction zone was dissociated due to the specific recognition of ATP with aptamer and released into the detection zone with the assistance of controllable fluid separator. The free G-quadruplex/hemin could catalyze hydrogen peroxide to generate oxygen for the consumption of photo-induced electrons from CuO NFs, which could further promote the electron-hole carriers separation efficiency, and eventually resulting in the enhancement of PEC signal. The proposed PEC lab-on-paper device could be employed for specific detection of ATP in the range from 5.0 to 3.0 × 103 nM with a detection limit of 2.1 nM.

Paper-Supported Self-Powered System Based on a Glucose/O2 Biofuel Cell for Visual MicroRNA-21 Sensing.

The exploitation of self-powered devices that get rid of the power source restriction represents the development tendency of sensing systems. Herein, a paper-supported glucose/O2 biofuel cell (BFC)-based self-powered sensing platform for visual analysis was developed. The BFC device utilized gold nanoparticle-modified paper fibers as the electrode to wire glucose oxidase (GOx) and bilirubin oxidase for the fabrication of bioanodes and biocathodes. To implement an assay protocol, a target-responsive cargo release system based on mesoporous silica nanocarriers controlled by microRNA-21 (miRNA-21) was designed. During the BFC operation, undesired H2O2, the side product of glucose oxidation which would be deleterious for GOx, was generated, leading to inevitable degeneration of BFC performance. On the basis of the H2O2-mediated iodide oxidation reaction to form iodine that further modulated the starch chromogenic reaction, undesired H2O2 could be effectively removed, resulting in remarkably improved BFC performance as well as providing a means for visual signal readout. Thanks to the dual output signals (maximum power output density or length of blue bar), enhanced analysis reliability and sensitive detection of miRNA-21 over a range of 5 fM to 100 pM were achieved. Moreover, this study demonstrates a proof of concept in visualized BFC-based self-powered systems for sensing applications and provides a blueprint to advance future sensors and analysis devices powered by BFCs in a wide variety of in vitro applications.