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Lung cancer is a major malignant cancer with low survival rates, and early diagnosis is crucial for effective treatment. Herein, a biosensing platform that is self-powered derived from a capacitor-coupled EBFC has been developed for ultra-sensitive real-time identification of microRNA-21 (miRNA-21) with the assistance of a mobile phone. The flexible substrate of the platform is prepared on a carbon paper modified with graphdiyne and gold nanoparticles. The biosensor employs DNAzyme-mediated dual strand displacement amplification, which enhances the signal output intensity of the EBFC and improves selectivity. The coupling of the capacitor with the EBFC significantly amplifies the sensing signal, causing a 10.6-fold surge in current respond and further improving the sensitivity of the sensing platform. The established detection approach demonstrates a linear relationship varied from 0.0001 to 10,000 pM, with a sensitivity down to 32.3 aM as the minimum detectable limit, which has been effectively utilized for detecting miRNA-21 in practical samples. This sensing system provides strong support for the construction of portable detection devices, and the strategy of the platform construction provides an effective method for ultra-sensitive and accurate detection of miRNA, holding great potential in clinical diagnosis, prognosis evaluation, and drug screening for cancer., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)

Carcinoembryonic antigen (CEA), a vital biomarker, plays a significant role in the early diagnosis and prognostic estimation of malignant tumors. In this study, a split-type photoelectrochemical immunoassay for the sensitive quantification of CEA has been successfully developed based on the target-induced in situ formation of a Z-type heterojunction. First, gold nanoparticle-decorated ZnIn 2 S 4 (AuNPs/ZnIn 2 S 4 ) composites were synthesized and used for the fabrication of photoelectrodes. Then, the detection antibody labeled with Ag nanoparticles was formed and applied for the biorecognition of CEA and subsequent liberation of Ag + ions to induce the in situ formation of Ag 2 S/AuNPs/ZnIn 2 S 4 , a Z-type heterojunction, on the photoelectrode. The Z-type Ag 2 S/AuNPs/ZnIn 2 S 4 heterojunction with effectively promoted separation of photogenerated charge carriers could lead to a markedly enhanced photocurrent response and highly sensitive quantification of CEA. Moreover, the three-dimensional spatial structure of ZnIn 2 S 4 provides abundant active sites for the reaction and exhibits non-enzymatic properties, which are conducive to the further improvement of the analytical performance of CEA. The developed split-type photoelectrochemical immunoassay with good sensitivity, satisfactory selectivity, reliable stability, wide dynamic linear range (0.01-20 ng mL -1 ), and low detection limit (7.3 pg mL -1 ) offers valuable insights into the development of novel PEC biosensing models for the detection of tumor biomarkers and holds potential application value in the field of disease diagnosis.

Gold nanoparticles (AuNPs), consisting of metallic gold, are applied in various fields owing to their characteristic physical properties. Collimonas sp. D-25 (D-25) is a Gram-negative bacterium obtained from soil, compost, and other environmental materials in the Akita Prefecture. DP-1 is a water-soluble protein found in D-25 that binds specifically to AuNPs and retains them with high stability. This study aimed to identify the part of DP-1 that interacts with AuNPs and determine its 3D structure in solution using nuclear magnetic resonance spectroscopy. Peptide fragments obtained by trypsin digestion were examined for their AuNP-binding capacity to determine the key Au-binding domain of DP-1. A fragment consisting of 16 amino acid residues (GHAATPEQYGVVTANK) was identified as the peptide with the highest binding activity. Structural analyses of this peptide indicated that the main chain was elongated, and negatively charged residues in the side chain were exposed on the surface by incorporating AuNPs. These results suggest that DP-1 interacts with AuNPs through negatively charged residues and extended hydrophobic residues for protein-protein interactions. The structural data also provide new insights into biomimetic technologies., (© 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.)

Based on the controllable instantaneous self-assembly ability of long-chain branched DNA nanostructures and the synergistic effect between nucleic acid amplification without enzymes, a highly sensitive and highly specific self-powered biosensing platform is developed. Two-dimensional graphdiyne is prepared, modified on flexible carbon cloth, and then functionalized with gold nanoparticles. When DNA mi-tubes are applied on it, target thalassemia gene CD122 triggers a dual-catalytic hairpin assembly reaction. The generated nanoscale DNA is precisely captured by the DNA mi-tube, exposing binding sites and activating the hybridization chain reaction to form long-chain branched DNA. Double-stranded DNA, along with dendritic DNA carrying a large number of guanine bases, precisely captures the signal molecule methylene blue (MB), generating a significant electrochemical signal. The redox reaction of MB also causes a proportional change in the system's color, achieving a colorimetric detection functionality. An efficient dual-mode self-powered sensing platform, therefore, is established for detecting the thalassemia gene CD122. The linear response range of target concentration to open-circuit voltage and RGB Blue value is 0.0001-10,000 pM. The detection limit under electrochemical mode is 36.3 aM ( S / N = 3), and under colorimetric mode, it is as low as 12.1 aM ( S / N = 3). The new method exhibits high sensitivity, excellent selectivity, and high accuracy, providing a universal strategy for designing novel biosensing platforms that can be extended to the detection of other biomolecules.

A new photoelectrochemical immunoassay based on self-assembled p-n Ag 2 O@Bi 2 O 2 S nanoflower heterojunction was designed and developed for quantitative monitoring of prostate-specific antigen (PSA) in biological fluids. Primarily, self-assembled p-n Ag 2 O@Bi 2 O 2 S nanoflower heterojunctions were served as the photoactive materials and coated onto the surface of electrodes. Subsequently, the glucose oxidase (GOx) was bound to the detection antibody (mAb 2 ) labeled gold nanoparticles (Au NPs) and then were employed to accomplish a sandwich-like immunoreaction to generate H 2 O 2 on a microplate incubated with monoclonal anti-PSA antibodies. In the presence of PSA, the product (H 2 O 2 ) was catalyzed by the substrate, which was used as an electron sacrificial agent to improve signal conversion and capture of photogenerated electrons. Under optimum conditions, a wide linear range of 0.01-50 ng mL -1 and a low detection limit of 5.3 pg mL -1 were accomplished with the sensor, exhibiting an excellent photocurrent response. Moreover, the proposed sensor revealed satisfactory reproducibility, high selectivity, and acceptable accuracy for the real sample testing. Importantly, our work provides a novel strategy for high sensitivity detection of disease-associated biomarkers for the early diagnosis of cancers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Photoelectrochemical (PEC) immunoassay has been widely developed for biomarker detection, but most include heavy and expensive instruments that are not suited for portable and on-site detection. In this work, the PEC immunoassay platform for mobile phones was reported for flexible, rapid, low-cost detection of carcinoembryonic antigen (CEA). The PEC detection platform was successfully composed of disposable screen-printed carbon electrodes, a micro-electrochemical workstation, a flashlight (the excitation light source), and a smartphone with a companion software with a micro-electrochemical workstation for rapid and on-site detection of target biomarkers. In this portable smartphone-based PEC system, the S-scheme heterojunction BiOCl/CuBi 2 O 4 was effectively excited due to the efficient electron transfer rate and excellent photocurrent response under visible light. Specifically, the sandwich-type immunoreaction for capturing target biomarkers introduced alkaline phosphatase (ALP) labeled gold nanoparticles (Au NPs). The addition of CEA increased the ascorbic acid (AA) content and enhanced the photocurrent. The proposed immunoassay presented a good linear with the logarithm of CEA concentrations range within 0.01-40 ng mL -1 , and the detection limit of 3.5 pg mL -1 (S/N = 3). Therefore, the portable detection platform offered an implementable approach to the development of miniaturized and portable photoelectrochemical detectors and on-site detection technology., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Thalassemia is one of the most common monogenic diseases, which seriously affects human growth and development, cardiovascular system, liver, etc. There is currently no effective cure for this disease, making screening for thalassemia particularly important. Herein, a self-powered portable device with high sensitivity and specificity for efficiently screening of low-level thalassemia is developed which is enabled with AuNPs/MoS 2 @C hollow nanorods and triple nucleic acid amplification technologies. It is noteworthy that AuNPs/MoS 2 @C electrode shows the advantages of high electrocatalytic activity, fast carrier migration rate and large specific surface area, which can significantly improve the stability and output signal of the platform. Using high-efficiency tetrahedral DNA as the probe, the target CD122 gene associated with thalassemia triggers a catalytic hairpin assembly reaction to achieve CD122 recycling while providing binding sites for subsequent hybridization chain reaction, greatly improving the detection accuracy and sensitivity of the device. A reliable electrochemical/colorimetric dual-mode assay for CD122 is then established, with a linear response range of 0.0001-100 pM for target concentration and open circuit voltage, and the detection limit is 78.7 aM (S/N = 3); a linear range of 0.0001-10000 pM for CD122 level and RGB Blue value, with a detection limit as low as 58.5 aM (S/N = 3). This method achieves ultra-sensitive and accurate detection of CD122, providing a new method for the rapid and accurate screening of thalassemia., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Collimonas sp. (D-25), found in the soil of Akita Prefecture, is a gram-negative bacterium with the ability to synthesize gold nanoparticles (AuNPs). During the synthesis of AuNPs, one specific protein (DP-1) was found to have disappeared in the sonicated solution of the bacterium. Recombinant DP-1 (rDP-1) from Escherichia coli BL21 (DE3) was used to study the effect of DP-1 on the synthesis of AuNPs. AuNPs synthesized with rDP-1 result in small, stabilized nanoparticles. AuNPs synthesized by DP-1 retained the stability of both the dispersion and nano-size particles under high salt concentrations. Isothermal titration calorimetry was employed to investigate the bonding ratio of rDP-1 to AuNPs. Several thousand rDP-1 proteins are attached to the surface of an AuNP to form a protein corona containing multiple layers. These results suggest that DP-1 obtained from D-25 has a size and stability control function during AuNP synthesis., (© 2023 The Authors. ChemBioChem published by Wiley-VCH GmbH.)

Herein, we presented a dual-readout gasochromic immunosensing platform for accurate and sensitive detection of carcinoembryonic antigen (CEA) based on Ag-doped/Pd nanoparticles loaded MoO 3 nanorods (Ag/MoO 3 -Pd). Initially, the presence of analyte CEA would prompt the formation of sandwich-type immunoreaction, accompanied by the introduction of Pt NPs labeled on detection antibody. Upon the addition of NH 3 BH 3 , the product hydrogen (H 2 ) will interact with Ag/MoO 3 -Pd as a bridge between the sensing interface and the biological assembly platform. Both photocurrent and temperature signals can serve as readouts due to the significantly increased PEC performance and enhanced photothermal conversion capability of H-Ag/MoO 3 -Pd (the product of Ag/MoO 3 -Pd react with H 2 ) compared to Ag/MoO 3 -Pd. In addition, the DFT results show that the band gap of Ag/MoO 3 -Pd becomes narrower after the reaction with H 2 , thus improving the utilization of light, which theoretically explains the internal mechanism of gas sensing reaction. Under optimal conditions, the designed immunosensing platform showed good sensitivity for CEA detection with the limit of detection (LOD) of 26 pg mL -1 (photoelectrochemical mode) and 98 pg mL -1 (photothermal mode). This work not only presents the possible reaction mechanism of Ag/MoO 3 -Pd and H 2 , but also creatively applicate it in photothermal biosensors that give a new path for devising dual-readout immunosensor., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 Elsevier B.V. All rights reserved.)

Accurate identification of acutely toxic and low-fatality mycotoxins on a large scale in a quick and cheap manner is critical for reducing population mortality. Herein, a portable photothermal immunosensing platform supported by a microelectromechanical microsystem (MEMS) without enzyme involvement was reported for point-of-care testing of mycotoxins (in the case of aflatoxin B 1 , AFB 1 ) in food based on the precise satellite structure of Au nanoparticles. The synthesized Au nanoparticles with a well-defined, graded satellite structure exhibited a significantly enhanced photothermal response and were coupled by AFB 1 antibodies to form signal conversion probes by physisorption for further target-promoted competitive responses in microplates. In addition, a coin-sized miniature NIR camera device was constructed for temperature acquisition during target testing based on advanced MEMS fabrication technology to address the limitation of expensive signal acquisition components of current photothermal sensors. The proposed MEMS readout-based microphotothermal test method provides excellent AFB 1 response in the range of 0.5-500 ng g -1 with detection limits as low as 0.27 ng g -1 . In addition, the main reasons for the efficient photothermal transduction efficiency of Au with different graded structures were analyzed by finite element simulations, providing theoretical guidance for the development of new Au-based photothermal agents. In conclusion, the proposed portable micro-photothermal test system offers great potential for point-of-care diagnostics for residents, which will continue to facilitate immediate food safety identification in resource-limited regions.

This work presents a photocurrent-polarity-switching-based photoelectrochemical (PEC) biosensing platform for ultrasensitive detection of microRNA-21 (miR-21) through target-triggered catalytic hairpin assembly (CHA) for modulation of methylene blue (MB) and ferrocene (Fc) positional configurations using double-shelled Cu-doped ZnS nanocages (NCs)-Au nanoparticles (NPs) as photoactive materials. In the presence of miR-21, the assembly of MB-labeled HP1 and Fc-labeled HP2 leads to the generation of a large amount of double-stranded DNA (HP1-HP2), which pushes MB away from the electrode surface and brings Fc close to the electrode surface, resulting in effectively quenching the enhanced PEC signal to activate the photocurrent-polarity-switching system. Benefiting from the distance-controllable strategy, the designed PEC bioanalysis can effectively eliminate false-positive and false-negative signals due to the change of different signal expression patterns (from traditional the "signal-on" mode to the photocurrent-polarity-switching mode), thereby significantly improving the sensing specificity and sensitivity. The proposed PEC sensing system exhibited satisfying photocurrent responses toward target miR-21 within the working range from 1.0 fM to 1 nM at a low limit of detection (LOD) of 0.58 fM. More importantly, we demonstrated the successful integration of the proposed PEC biosensor with a handheld wireless device for instant detection of miR-21 concentrations in practical samples.

Methods derived from photoelectrochemical (PEC) have been constructed for immunoassays, but most involve the split-type immunoreaction modes, and thus easily cause unpredictable intermediate precision. Herein, we innovatively designed an integrated PEC immunosensing platform for the quantitative monitoring of thyroglobulin (TG) on the gold nanoparticles (AuNPs)-functionalized BiVO 4 photoanode coupling with enzymatic biocatalytic precipitation (EBCP). This sensing system could simultaneously implement the immunoreaction and photocurrent measurement. Anti-TG capture antibodies were modified onto AuNPs-decorated BiVO 4 photoelectrode. A sandwich-type immunoreaction was carried out in the presence of target TG using horseradish peroxidase (HRP)-conjugated anti-TG detection antibody. The carried HRP molecules catalyzed 4-chloro-1-naphthol (4-CN) to generate an insoluble benzo-4-chlorohexadienone product on the photoanode in the presence of peroxide hydrogen, thereby decreasing the photocurrent. Under optimal conditions, the PEC immunosensors gave good photocurrent responses toward target TG within the dynamic range of 0.01-10 ng mL -1 at a detection limit of 7.6 pg mL -1 . Good repeatability and precision, high specificity and acceptable storage stability were acquired during the measurement. No significant differences were encountered for screening 15 human serum specimens between the developed PEC immunoassay and commercially available enzyme-linked immunosorbent assay (ELISA) method for the detection of target TG. Significantly, PEC immunosensing system offers promise for simple and cost-effective analysis of disease-related biomarkers., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Large-scale, rapid, and inexpensive serological diagnoses of severe acute respiratory syndrome-coronavirus-2 (SARS-CoV-2) are of great interest in reducing virus transmission at the population level; however, their development is greatly plagued by the lack of available point-of-care methods, leading to low detection efficiency. Herein, an ultrasensitive smartphone-based electrochemical immunoassay is reported for rapid (less than 5 min), low-cost, easy-to-implement detection of the SARS-CoV-2 nucleocapsid protein (SARS-CoV-2 N protein). Specifically, the electrochemical immunoassay was fabricated on a screen-printed carbon electrode coated with electrodeposited gold nanoparticles, followed by incubation of anti-N antibody (Ab) and bovine serum albumin as the working electrode. Accompanied by the antigen-antibody reaction between the SARS-CoV-2 N protein and the Ab, the electron transfer between the electroactive species [Fe(CN) 6 ] 3- / 4- and the electrode surface is disturbed, resulting in reduced square-wave voltammetry currents at 0.075 V versus the Ag/AgCl reference electrode. The proposed immunoassay provided a good linear range with SARS-CoV-2 N protein concentrations within the scope of 0.01-1000 ng/mL (R 2 = 0.9992) and the limit of detection down to 2.6 pg/mL. Moreover, the detection data are wirelessly transmitted to the interface of the smartphone, and the corresponding SARS-CoV-2 N protein concentration value is calculated and displayed. Therefore, the proposed portable detection mode offers great potential for self-differential diagnosis of residents, which will greatly facilitate the effective control and large-scale screening of virus transmission in resource-limited areas.

The point-of-care (POC) method with affordability and portability for the sensitive detection of biological substances is an emerging topic in rapid disease screening and personalized medicine. In this work, we demonstrated a diverse responsive platform based on a dual-channel pressure sensor (DCPS). The DCPS had a multilayer flexible architecture consisting of a photonic hydrogel with chromatic transitions and a piezoresistive pressure sensor as the electrical data transmission unit, both of which had the property of pressure-induced mechanical stimulus feedback. By incorporating a platinum nanoparticles-labeled detection antibody (PtNPs-dAb) into the sandwich-type immunoreaction for the target carcinoembryonic antigen (CEA, as a model analyte), gas decomposition could be triggered by the addition of hydrogen peroxide (H 2 O 2 ) to induce a significant increase under pressure in a closed chamber. Meanwhile, the DCPS enabled an accurate electrical signal output, and the photonic hydrogel converted spatiotemporal stimuli into eye-readable colorations with string brilliance. In this way, the target concentration could be quantificationally related to the electrical response and intuitively perceived through visible color alterations. Under optimal conditions, a sensitive determination of CEA was performed in a detectable range of 0.3-60 ng/mL with a limit of detection (LOD) of 0.13 ng/mL. In addition, the proposed protocol had satisfactory selectivity, accuracy, and reproducibility. Furthermore, an array-based immunoassay device was fabricated to conceptually validate its application potential in high-throughput biomedical detection and inspire a dual-signal POC diagnostic platform in a friendly way for resource-limited settings.

Functional photothermal nanomaterials have gained widespread attention in the field of precise cancer therapy and early disease diagnosis due to their unique photothermal conversion properties. However, the relatively narrow temperature response range and the outputable accuracy of commercial thermometers limit the accurate detection of biomarkers. Herein, we designed a liposome-embedded Cu 2- x Ag x S amplification-based photothermal sensor for the accurate determination of cardiac troponin I (cTnI) in health monitoring and point-of-care testing (POCT). The combinable 3D-printing detecting device monitored and visualized target signal changes in the testing system under the excitation of near-infrared (NIR) light, which was recorded and evaluated for possible pathogenicity by a smartphone. Notably, we predicted the potentially efficient thermal conversion efficiency of Cu 2- x Ag x S from the structure and charge density distribution, calculated by the first-principles and density functional theory (DFT), which provided a theoretical basis for the construction of novel photothermal materials, and the experimental results proved the correctness of the theoretical projections. Under optimal conditions, the photothermal immunoassay showed a dynamic linear range of 0.02-10 ng mL -1 with a detection limit of 11.2 pg mL -1 . This work instructively introduces promising theoretical research and provides new insights for the development of sensitive portable photothermal biosensors.

Ultra-sensitive detection of target molecules by surface-enhanced Raman scattering (SERS) is crucial in a wide range of fields but remains a great challenge. In this work, we report a simple and effective protocol for obtaining highly SERS-sensitive probe by mixing iodide with silver sol. The specific iodide effect on the SERS sensitivity is systematically investigated. It is found that, iodide can effectively promote the SERS enhancement of anionic and cationic analytes, and I - ion has a higher activating effect on SERS than that of Cl - ion. The as-prepared SERS-active substrate demonstrates excellent enhancement for rhodamine 6G with a high Raman enhancement factor of 1.8 × 10 8 , which allows the detection limit of 1.0 × 10 -13 M. Our findings in this work should be important for the developing of SERS theory and ultra-sensitive detection applications., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 Elsevier B.V. All rights reserved.)

Ligand-functionalized plasmonic nanoparticles have been widely used for targeted imaging in living systems. However, ligand presentation and encoding on the nanoparticle's surface in a stoichiometrically controllable manner remains a great challenge. Herein, we propose a method to construct ligand-engineered plasmonic nanoprobes by using nanoparticle encapsulation with topological DNA tetrahedrons, which enables the programmed ligand loading for precise regulation of targeting efficiency of nanoprobes in biorelated applications. With this method, we demonstrated the preparation of functionalized plasmonic nanoprobes by programmed loading of RGD peptides and aptamers onto the DNA tetrahedron encapsulated gold nanoparticles with controllable stoichiometric ratios. The cell imaging and particle counting assays suggested that the targeting efficiency of the nanoprobes could be readily modulated by tailoring the number and stoichiometric ratios of the loaded ligands, respectively. It can be anticipated that this robust strategy could provide new opportunities for the construction of efficacious nanoprobes and delivery systems for versatile bioapplications.

Early diagnosis of cancers relies on the sensitive detection of specific biomarkers, but most of the current testing methods are inaccessible to home healthcare due to cumbersome steps, prolonged testing time, and utilization of toxic and hazardous substances. Herein, we developed a portable self-powered photoelectrochemical (PEC) sensing platform for rapid detection of prostate-specific antigen (PSA, as a model disease-related protein) by integrating a self-powered photoelectric signal output system catalyzed with chemiluminescence-functionalized Au nanoparticles (AuNPs) and a phosphomolybdic acid (PMA)-based photochromic visualization platform. TiO 2 -g-C 3 N 4 -PMA photosensitive materials were first synthesized and functionalized on a sensor chip. The sensor consisted of filter paper modified with a photocatalytic material and a regional laser-etched FTO electrode as an alternative to a conventional PEC sensor with a glass-based electrode. The targeting system involved a monoclonal anti-PSA capture antibody-functionalized Fe 3 O 4 magnetic bead (mAb 1 -MB) and a polyclonal anti-PSA antibody (pAb 2 )- N -(4-aminobutyl)- N -ethylisoluminol-AuNP (ABEI-AuNP). Based on the signal intensity of the chemiluminescent system, the photochromic device color changed from light yellow to heteropoly blue through the PMA photoelectric materials integrated into the electrode for visualization of the signal output. In addition, the electrical signal in the PEC system was amplified by a sandwich-type capacitor and readout on a handheld digital multimeter. Under optimum conditions, the sensor exhibited high sensitivity relative to PSA in the range of 0.01-50 ng mL -1 with a low detection limit of 6.25 pg mL -1 . The flow-through chemiluminescence reactor with a semiautomatic injection device and magnetic separation was avoid of unstable light source intensity inherent in the chemiluminescence process. Therefore, our strategy provides a new horizon for point-of-care analysis and rapid cost-effective clinical diagnosis.

Prostate-specific antigen (PSA), a glycoprotein that is most likely to cause prostate cancer, has attracted widespread attention in recent years due to its increasing threat to people's lives and health. Herein, we developed a new signal-amplified photoelectrochemical (PEC) immunosensing method for quantitative monitoring of the target PSA based on the ion-exchange reaction for the in situ formation of ZnO/CdS/Ag 2 S nanohybrids triggered by the as-released silver ions (Ag + ) from silver nanolabels. Initially, the introduction of a target PSA caused the formation of a sandwich immunocomplex in an anti-PSA capture antibody (cAb)-coated microplate with the help of a silver nanoparticle-labeled detection antibody (AgNPs-dAb). Thereafter, the introduced AgNPs were dissolved with acid to release numerous silver ions. In this regard, an ion-exchange reaction occurred between the silver ions and ZnO/CdS nanorods on the photosensitive electrode, thus producing ZnO/CdS/Ag 2 S nanohybrids to generate a relatively strong photocurrent. Under optimal conditions, the ion-exchange reaction-based PEC immunoassay exhibited a good linear range of 0.05-50 ng mL -1 and allowed the detection of the target PSA at a concentration as low as 0.018 ng mL -1 . In addition, the PEC immunoassay displayed satisfactory repeatability, high specificity, and acceptable method accuracy. Importantly, the ion-exchange reaction-based PEC immunoassay provides a new perspective for the detection of other disease-related biomarkers by controlling the corresponding antibodies.

The in situ glycan profiling of a single tumor cell plays an important role in personalized cancer treatment. Herein, an integrated microfluidic system was designed for living single-cell trapping and real-time monitoring of galactosyl expression on the surface, combining closed bipolar electrode (BPE) arrays and electrofluorochromic (EFC) imaging. Galactosyl groups on human liver cancer HepG2 cells were used as the model analysts, galactose oxidase (GAO) could selectively oxidize hydroxyl sites of galactosyl groups on the cell surface to aldehydes, and then biotin hydrazide (BH) was used to label the aldehydes by aniline-catalyzed hydrazone ligation. With the biotin-avidin system, nanoprobes were finally introduced to the galactosyl groups on the cell surface with avidin as a bridge, which was prepared by simultaneously assembling ferrocene-DNA (Fc-DNA) and biotin-DNA (Bio-DNA) on gold nanoparticles (AuNPs) due to their large surface area and excellent electrical conductivity. After a labeled single cell was captured in the anodic microchannel, the Fc groups attached on the cell surface were oxidized under suitable potential, and the nonfluorescent resazurin on the cathode was correspondingly reduced to produce highly fluorescent resorufin, collected by fluorescence confocal microscope. The combination of EFC imaging and BPE realized monitoring galactosyl group expression of 5.0 × 10 8 molecules per cell. Furthermore, the proposed platform had the ability to distinguish a single cancer cell from a normal cell according to the expression level of galactosyl groups and to dynamically monitor the galactosyl group variation on the cell surface, providing a simple and accessible method for the single-cell analysis.

This work demonstrated a pressure-based biosensor integrated with a flexible pressure sensor and an electrochromic device for visual detection. Initially, a sandwich-type immunoreaction for target carcinoembryonic antigen (CEA, as a model analyte) was carried out using the capture antibody (cAb) and platinum nanoparticles-labeled detection antibody (PtNPs-dAb) in a reaction cell. The added hydrogen peroxide (H 2 O 2 ) could be catalyzed by the PtNPs to generate oxygen (O 2 ). In a sealed chamber, the pressure increased with the overflowing O 2 . Meanwhile, a skin-inspired flexible pressure sensor with excellent sensing performance was fabricated to monitor the pressure change in real time. Thus, the electrical signal of the pressure sensor could reveal the target concentration. Moreover, a voltage-regulated electrochromic device based on polyaniline (PANI) and tungsten oxide (WO 3 ) was integrated into the platform to provide a visualized readout. According to the electrical signal of the pressure sensor, the electrochromic device would change its color from green to blue, which also revealed the target concentration and could be observed by the naked eye. Under optimal conditions, the biosensor presented a high sensitivity for CEA in a detectable range of 0.2-50 ng/mL. The limit of detection (LOD) was 94 pg/mL. The selectivity, reproducibility, and accuracy were also satisfying. Furthermore, this immunoassay gives a path for developing visualized biosensors in point-of-care settings.

This work reports the proof-of-concept of novel and powerful enzyme-free voltammetric aptasensor of colorectal cancer-related carcinoembryonic antigen (CEA) without the participation of any antibodies. The aptasensor was prepared by means of immobilizing thiolated CEA aptasensor on gold nanoparticle-deposited glassy carbon electrode. Thionine-doped silica nanoparticles were synthesized via reverse micelle method and functionalized with p-aminophenylboronic acid by the epoxy-amino reaction for specific conjugation of subsequent CEA glycoprotein. In the presence of target CEA, the analyte initially reacted with immobilized aptamer on the electrode to generate an aptamer-CEA complex. Then, phenylboronic acid immobilized on silica nanoparticle specifically recognized with CEA glycoprotein based on sugar-boronic acid interaction to form a sandwich-type complex on the electrode. Within the applied potentials, the doped thionine molecules into the silica nanoparticles were used as the electron mediators to produce well-defined voltammetric signals (vs. Ag/AgCl). Under optimum conditions, the voltammetric peak current increased linearly within the CEA concentration in the range from 1.0 pg mL -1 to 10 ng mL -1 , and the limit of detection was 0.49 pg mL -1 at 3δ. A repeatability and intermediate precision of ≤8.7% was accomplished at CEA levels in the batch-to-batch mode. The method was highly specific for CEA over other disease-related biomarkers. The accuracy and inter-laboratory validation of this system were evaluated for target CEA detection in human serum specimens, and the results were in accordance with those obtained by a commercial ELISA., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Methods based on enzyme labels or nano labels have been developed for immunoassays, but most of them have low sensitivity and are unsuitable for low-abundance protein diagnostics and biosecurity. Herein, an innovative quartz crystal microbalance (QCM) immunosensing method was designed for high-efficiency detection of carcinoembryonic antigen (CEA) from serum samples with colorectal cancer patients by using horseradish peroxidase (HRP) nanoparticles as the enhancer, accompanying enzymatic biocatalytic precipitation (EBCP) toward 4-chloro-1-naphthol (4-CN) on an anti-CEA capture antibody-conjugated QCM probe. Initially, HRP nanospheres were synthesized on the basis of the reverse micelle method with the assistance of glutaraldehyde cross-linking, and then covalently conjugated onto polyclonal anti-CEA detection antibodies. The QCM immunosensing probe was prepared by immobilizing monoclonal anti-CEA capture antibodies on an l-cysteine-modified gold substrate. In the presence of target CEA, a sandwich-type immunoreaction was carried out between capture antibody and detection antibody, thereby resulting in the attachment of HRP nanospheres on the gold probe. Upon addition of 4-CN in the QCM cell, the carried HRP nanospheres catalyzed the 4-CN oxidation to produce an insoluble precipitate on the gold electrode, thus causing a change in the frequency. Relative to the conventional HRP-labeled strategy and direct antigen-antibody reaction, improved analytical features were obtained with HRP nanospheres. With the HRP nanosphere labeling method, the factors influencing the performance of the immunoassay were also studied. The covalent conjugation of antibodies with HRP nanospheres and the gold substrate achieved a good repeatability and intermediate precision down to 10.7%. Under optimum conditions, the frequency variation of the QCM immunoassay was proportional to the target CEA concentration within a dynamic linear range of 0.01-100 ng mL-1 with a detection limit (LOD) of 7.8 pg mL-1. In addition, the developed QCM immunoassay showed high specificity and long-term storage stability, and could be used for the analysis of human serum samples with consistent results in comparison with those obtained from the commercial enzyme-linked immunosorbent assay (ELISA) method.

A paper-based visual fluorescence immunoassay is presented for the detection of matrix metalloproteinase-7 (MMP7) that is related to renal cancer. The method is based on the distance-dependent fluorescence quenching of CdTe quantum dots (QDs) on a nitrocellulose membrane by Ag + following a sandwich-type immunoreaction on microtiter wells using silver nanoparticle (AgNP)-labeled secondary antibody- and primary antibody-coated microtiter wells. The silver nanoparticles captured in the well are dissolved with HNO 3 , while the quenching effect of QDs is based on silver ion-exchange reaction under 365-nm excitation light irradiation. Increasing concentration of released Ag + , thus higher concentration of the protein, leads to an increased distance of quenching on the nitrocellulose membrane. The paper-based immunoassay by combination of AgNP-assisted ion-exchange reaction with QD gives good distance-dependent responses and allows the detection of MMP7 at a concentration as low as 7.3 pg mL -1 . The coefficients of variation are less than 6.9% and 12.4% for intra-assay and inter-assay, respectively. High specificity and long-term stability are achieved during the assay. Importantly, the testing of human serum samples using our strategy shows well-matched results with commercial human MMP7 ELISA kits. Graphical abstract A distance-dependent visual immunoassay is developed for the determination of serum matrix metalloproteinase-7 on CdTe quantum dot-impregnated paper with silver ion-exchange reaction.

This work describes a novel and portable pressure-based point-of-care (POC) testing strategy for the sensitive and rapid detection of carcinoembryonic antigen (CEA) via a flexible pressure sensor constructed by three-dimensional (3D) polypyrrole (PPy) foam. Initially, platinum nanoparticles (PtNPs) were conjugated to the detection antibodies, which were used to form sandwich-type immunocomplexes with targets and capture antibodies in the reaction cell. Then, the carried PtNPs catalyzed the dissociation of hydrogen peroxide (H 2 O 2 ) for the generation of oxygen (O 2 ) in a sealed device, translating the biomolecule recognition event into gas pressure. With the increase of pressure, a flexible pressure sensor with 3D polypyrrole foam as the sensing layer was used to sensitively monitor the pressure variations in this system. Thus, the concentration of the target could be quantitatively determined by the pressure response. Under optimal conditions, the pressure-based immunosensor showed good sensing performance for CEA in the dynamic working range from 0.2 to 60 ng/mL with a detection limit of 0.13 ng/mL. The reproducibility, specificity, and accuracy compared with commercial enzyme-linked immunosorbent assay (ELISA) kit were also acceptable. Therefore, this work provides a promising approach for developing portable and sensitive POC testing in the future.

A photoelectrochemical (PEC) aptasensing platform is devised for sensitive detection of an organophosphorus pesticide based on dissolution of core-shell MnO 2 nanoflower@CdS (MnO 2 NF@CdS) by thiocholine (TCh). TCH is produced from the butyrylcholinesterase-acetylthiocholine system, accompanied by target-triggered rolling circle amplification (RCA). The core-shell MnO 2 NF@CdS with excellent PEC performance was synthesized and employed as a photo-sensing platform. The target was detected on a functionalized magnetic probe with the corresponding aptamer. Upon malathion introduction, the aptamer was detached from the magnetic beads, while capture DNA (cDNA, with primer fragment) remained on the beads. The primer fragment in cDNA can trigger the RCA reaction to form a long single-stranded DNA (ssDNA). Furthermore, a large number of butyrylcholinesterase (BChE) were assembled on the long ssDNA strands through the hybridization with the S 2 -Au-BChE probe. Thereafter, TCh generated from hydrolysis of ATCh by BChE can reduce MnO 2 NF (core) to Mn 2+ and release the CdS nanoparticles (shell) from the platform electrode, significantly enhancing the PEC signal. Under optimal conditions, the proposed aptasensor exhibited high sensitivity for malathion with a low detection limit of 0.68 pg mL -1 . Meanwhile, it also presents outstanding specificity, reproducibility, and stability. Importantly, the sensing platform provides a new concept for detection of pesticide. Graphical abstract Herein, this work devised a photoelectrochemical (PEC) aptasensing platform for sensitive detection of organophosphorus pesticide based on dissolution of core-shell MnO 2 nanoflower@CdS (MnO 2 NF@CdS) by the as-produced thiocholine (TCh) from the butyrylcholinesterase-acetylthiocholine system, accompanying with the target-triggered rolling circle amplification (RCA).

The variations in microRNA (miRNA) expression levels can be useful biomarkers for the diagnosis of different cancers. In this work, a label-free and sensitive fluorescent method for detection of miRNA-21 is described based on duplex-specific nuclease (DSN) assist target recycling and terminal deoxynucleotidyl transferase (TdT) induced copper nanoclusters (CuNCs). In the absence of target, the 3'-phosphorylated probe DNA cannot be hydrolyzed by DSN and extended by TdT, and failed to synthesizing fluorescent CuNCs. However, the target miRNA-21 can caused the digestion of probe DNA with DSN, releasing primer DNA with 3'-OH. After that, the primer DNA can forms long poly T with the assistance of TdT, leading to synthesize high fluorescent CuNCs. The fluorescence change of CuNCs can be used to identify the concentration of target miRNA-21. Under optimal experimental conditions, this strategy could quantitatively detect miRNA-21 down to 18.7 pM. We have also demonstrated the practical application of our proposed method for monitoring miRNA-21 expression levels in cancer cells. Moreover, this method show good specificity for miRNA-21 detection due to the strong preference of DSN for cutting perfectly matched DNA/RNA duplex, which holds great potential for highly specific quantification of biomarkers in bioanalysis and clinical diagnosis., (Copyright © 2019 Elsevier Inc. All rights reserved.)

An innovative visible light-driven photoelectrochemical (PEC) immunosensing system was reasonably established for the sensitive detection of prostate-specific antigen (PSA) by using perovskite metal oxide@gold nanoparticle heterostructures (BaTiO 3 /Au) as the photoactive materials. When plasmonic Au nanoparticles were directly decorated on BaTiO 3 , a several times surface plasmon resonance (SPR) enhancement of photocurrent density was induced via the injection of hot electrons from visible light-excited Au nanoparticles into the conduction band of BaTiO 3 , and the combination of BaTiO 3 and Au nanoparticles was employed as a promising platform for developing a photoelectrochemical bioanalysis. As a proof of concept, PSA had been detected by the BaTiO 3 /Au nanocomposite-based PEC sensor. To design such an immunoassay protocol, a monoclonal anti-PSA capture antibody (cAb)-coated microplate and glucose oxidase/polyclonal anti-PSA detection antibody-modified gold nanoparticles (GOx-Au NP-dAb) were used as the immunoreaction platform and signal probe, respectively. Upon the addition of target PSA, a sandwiched immunocomplex was formed accompanying the immuno-recognition between the antigen and antibody, and then the carried GOx could oxidize glucose to produce H 2 O 2 . The photocurrent of the BaTiO 3 /Au nanocomposite-functionalized electrode amplified with increasing H 2 O 2 concentration since H 2 O 2 is considered as a good hole scavenger. On the basis of the above-mentioned mechanisms and the optimized conditions, the assembled PEC immunosensor was linear with the logarithm of the PSA concentration in the range of 0.01-40 ng mL -1 with a detection limit of 4.2 pg mL -1 . It afforded rapid response, good precision, and high stability and specificity, implying its great promise in photoelectrochemical immunoassays. More generally, this system sets up an ideal PEC immunosensing system based on the BaTiO 3 /Au nanocomposites and represents an innovative and low-cost "signal-on" assay scheme for the practical quantitative screening of low-abundance proteins.

Developing a theranostic platform that integrates diagnosis and treatment in one single nanostructure is necessary for efficient tumor treatment. Here, we presented a novel theranostic nanoprobe for nonlabeled fluorescence imaging of Zn 2+ and 635 nm red light-triggered photodynamic therapy (PDT) by a multifunctional DNA-templated silver nanocluster/porphyrin/MnO 2 nanoplatform. MnO 2 nanosheets adsorbed hairpin DNA-silver nanoclusters (AgNCs) and porphyrin (P) by facile physisorption, which accelerate the transfection of nanoprobes and P into tumor cells. After entering the cells, the biodegradation of MnO 2 nanosheets by glutathione and acidic hydrogen peroxide released AgNCs for label-free Zn 2+ fluorescence imaging by the hairpin DNA-fueled dynamic self-assembly of three-way DNA junction architectures, and the released Mn 2+ could act as an effective magnetic resonance imaging (MRI) contrast agent. In addition, MnO 2 was decomposed in the acidic H 2 O 2 -ample environment and produced O 2 to overbear hypoxia-related PDT resistance, highly efficient PDT was obtained by excess singlet oxygen ( 1 O 2 ) release of P-AgNCs-MnO 2 nanoprobes under light irradiation compared with free P. In vitro and in vivo studies confirmed that P-AgNCs-MnO 2 exhibited high fluorescence specificity, excellent PDT effect, and good biocompatibility and could be used as a contrast agent for MRI. This theranostic platform provided a new avenue for the fluorescence and MRI diagnosis of tumors and efficient tumor treatment.

This work presented an innovative and rationally engineered palindromic molecular beacon (PMB) based "Z-scheme" photoelectrochemical (PEC) biosensing protocol for the selective screening of kanamycin (Kana) through DNA hybridization-induced conformational conversion. Interestingly, the ingeniously designed PMB integrated the multifunctional elements including recognition region, primer-like palindromic fragment, and polymerization-nicking template. The cosensitized structures consisted of CdS quantum dot functionalized hairpin DNA2 (QD-HP2) and region-selectively deposited gold nanoparticles onto {001} facets of bismuth oxychloride (BiOCl-Au). Compared with BiOCl-Au alone, the attachment of CdS QDs onto BiOCl-Au (i.e., BiOCl-Au-CdS QDs) exhibited evidently enhanced photocurrent intensity thanks to the synergistic effect of Z-scheme BiOCl-Au-CdS QDs. After incubation with target Kana, Kana-aptamer binding could induce the exposure of PMB region for hairpin DNA1 (HP1). The exposed palindromic tails hybridized with each other (like a molecular machine) to consume the substrates (dNTPs) and fuels (enzyme) for the releasing of numerous nick fragments (Nick). The as-generated nick fragments could specifically hybridize with the complementary region of QD-HP2, thus resulting in decreasing photocurrent because of the increasing spatial distance for electron transfer between two-type photosensitizers. Under optimum conditions, the PMB-based sensing system exhibited satisfying photocurrent responses toward target Kana within the working range from 50 to 5000 fM at a low detection limit of 29 fM. Impressively, the concept of a palindromic fragment-mediated primer-free biosensing strategy offers a new avenue for advanced development of efficient and convenient biodetection systems.

A multifunctional nanoplatform that enables the integration of biological detection, imaging diagnosis, and synergistic therapy into a single nanostructure holds great promise for nanoscience and nanomedicine. Herein, a novel theranostic platform was presented for label-free imaging of cell surface glycans based on DNA/silver nanoclusters (AgNCs) via hybridization chain reaction (HCR) and fluorescence guided photothermal therapy (PTT). In this strategy, a dibenzocyclooctyne (DBCO)-functionalized DNA and two hairpin structures of DNA/AgNCs probes were involved. Following metabolic glycan labeling, the binding of DBCO-functionalized DNA to cell surface initiated HCR, and then cell surface glycans were specifically labeled by DNA/AgNCs fluorescent probes. Furthermore, this signal amplification strategy was adopted in quantitative analysis, and the detection limit could be achieved as low as 20 cells in 200 μL of binding buffer. Moreover, the remarkable photothermal properties of DNA/AgNCs via HCR led to efficient killing of cancer cells and inhibited the tumor growth under imaging guide. In this strategy, DNA/AgNCs were utilized to detect the cellular glycans, which aided in overcoming the high cost and instability of fluorescent dyes. Simultaneously, the HCR process avoided the introduction of excessive azido-sugars under the precondition of ensuring apparent fluorescence. These results indicated that the developed nanoplatform has great potential for specific cell surface glycans imaging and fluorescence guided PTT.

A competitive colorimetric assay has been established to detect chloramphenicol (CAP). It is based on the use of colloidal and electrostatically stabilized aptamer-modified gold nanoparticles (GNPs). The CAP aptamer is modified by a sequence of 5 adenosine groups to anchor it on the surface of GNPs. It can competitively capture two compounds, viz. D-(-)-threo-2-amino-1-(4-nitrophenyl)-1,3-propanediol (CAP-base, with a positive charge) and CAP (which is uncharged). The capture of the positively charged CAP-base triggers the aggregation of modified GNPs in salt-containing solution, and this causes a color change from red to purple. However, in the presence of CAP and CAP-base, the capture of the uncharged CAP weakens this color change by a competing process for capture. Thus, the concentration of CAP is associated with the degree of deaggregation of GNPs and can be quantified by the ratio of absorbances at 620 nm and 520 nm. The assay has a 22 nM limit of detection in acidic solution, and the response is linear in the range of 0.20 to 3.20 μM CAP concentration. This assay was successfully applied to the determination of CAP in spiked environmental water samples. Conceivably, this method has a wide scope in that it may be applied to a wide range of analytes if respective aptamers are available. Graphical abstract Schematic presentation of a competitive non-cross linking deaggregating method for detecting chloramphenicol. The surface charge of polyA-Apt@GNPs and its aggregation degree (purple) are determined by the charge of target. (CAP-base: precursor of CAP; PolyA-Apt@GNPs: 5'-polyA-modified DNA aptamer functionalized gold nanoparticles.).

A new photoluminescence (PL) enzyme immunoassay was designed for sensitive detection of aflatoxin B 1 (AFB 1 ) via an innovative enzyme substrate, 6-aza-2-thiothymine-stabilized gold nanocluster (AAT-AuNC) with L-arginine. The enzyme substrate with strong PL intensity was formed through supramolecular host-guest assembly between guanidine group of L-arginine and AAT capped on the surface of AuNC. Upon arginase introduction, the captured L-arginine was hydrolyzed into ornithine and urea, thus resulting in the decreasing PL intensity. Based on this principle, a novel competitive-type immunoreaction was first carried out on AFB 1 -bovine serum albumin (AFB 1 -BSA) conjugate-coated microplate, using arginase-labeled anti-AFB 1 antibody as the competitor. Under the optimum conditions, the PL intensity increased with the increment of target AFB 1 , and allowed the detection of the analyte at concentrations as low as 3.2 pg mL -1 (ppt). Moreover, L-arginine-AAT-AuNC-based PL enzyme immunoassay afforded good reproducibility and acceptable specificity. In addition, the accuracy of this methodology, referring to commercial AFB 1 ELISA kit, was evaluated to analyze naturally contaminated or spiked peanut samples, giving well-matched results between two methods, thus representing a useful scheme for practical application in quantitative monitoring of mycotoxins in foodstuff., (Copyright © 2018 Elsevier B.V. All rights reserved.)

A competitive-displacement reaction strategy based on target-induced dissociation of gold nanoparticle coated graphene nanosheet (AuNPs/GN) from CdS quantum dot functionalized mesoporous titanium dioxide (CdS QDs/TiO 2 ) was designed for the sensitive photoelectrochemical (PEC) aptasensing of prostate-specific antigen (PSA) through the exciton-plasmon interaction (EPI) between CdS QDs and AuNPs. To construct such an aptasensing system, capture DNA was initially conjugated covalently onto CdS QDs/TiO 2 -modified electrode, and then AuNPs/GN-labeled PSA aptamer was bound onto biofunctionalized CdS QDs/TiO 2 via hybridization chain reaction of partial bases with capture DNA. Introduction of AuNPs/GN efficiently quenched the photocurrent of CdS QDs/TiO 2 thanks to energy transfer. Upon addition of target PSA, the sandwiched aptamer between CdS QDs/TiO 2 and AuNPs/GN reacted with the analyte analyte, thus resulting in the dissociation of AuNPs/GN from the CdS QDs/TiO 2 to increase the photocurrent. Under optimum conditions, the aptasensing platform exhibited a high sensitivity for PSA detection within a dynamic linear range of 1.0 pg/mL to 8.0 ng/mL at a low limitat of detection of 0.52 pg/mL. The interparticle distance of exciton-plasmon interaction and contents of AuNPs corresponding to EPI effect in this system were also studied. Good selectivity and high reproducibility were obtained for the analysis of target PSA. Importantly, the accuracy and matrix effect of PEC aptasensor was evaluated for the determination of human serum specimens and newborn calf serum-diluted PSA standards, giving a well-matched result with the referenced PSA ELISA kit.

The authors describe a fluorometric strategy for the detection of pathogenic bacteria with ultrasensitivity and high specificity. This strategy relies on the combination of target-modulated photoinduced electron transfer (PET) between G-quadruplex DNAzyme and DNA (labeled with silver nanoclusters) along with hairpin probe-based circular exponential amplification. The reaction system involves three hairpin probes (H1, H2 and H3). Probe H1 contains an aptamer against S. Typhimurium and the recognition sequence for nicking endonuclease. It is used to recognize S. Typhimurium and participates in polymerase-catalyzed target recycle amplification and secondary-target recycle amplification. Probe H2 contains an aptamer against hemin and is used to form the G-quadruplex DNAzyme in the presence of hemin and potassium ion. It acts as the electron acceptor and quenches the fluorescence of the labeled DNA. Fluorescence is best measured at excitation/emission wavelengths of 567/650 nm. Probe H3 contains the template sequence for the synthesis of AgNCs and the H2-annealing sequence. Both H2 and H3 are utilized to perform a strand displacement reaction and to achieve PET between G-quadruplex DNAzyme and DNA/AgNCs. To the best of our knowledge, this is the first example of a PET between G-quadruplex DNAzyme and DNA/AgNCs coupled with circular exponential amplification. The assay has an ultra-low detection limit 8 cfu·mL -1 of S. Typhimurium. The assay is rapid, accurate, inexpensive and simple. Hence, the strategy may represent a useful platform for ultrasensitive and highly specific detection of pathogenic bacteria as encountered in food analysis and clinical diagnosis. Graphical abstract The reaction system includes three hairpin probes (H1, H2 and H3), primer probe (P), Phi 29 DNA ploymerase (Phi 29) and nicking endonuclease Nt.AlwI (Nt.AlwI). Phi 29 and Nt.AlwI -assisted signal amplification leads to the recycling of target and produces numerous single stranded-DNAs (S). Strand displacement amplification leads to photoinduced electron transfer (PET) between G-quadruplex DNAzyme and DNA/AgNCs. HAP-based circular exponential amplification of PET results in an ultrasensitive fluorometric assay.

Herein, a simple and feasible electrochemical immunosensing method for simultaneous voltammetric detection of two immunoglobulin proteins, human IgG (HIgG) and rabbit IgG (RIgG), was developed using two distinguishable signal-generation tags on the same electrode. The immunosensor was prepared by immobilizing two Fab antibody fragments on a gold electrode. After this, Cu and Cd nanocrystals, as nanotags, were synthesized and functionalized with identical detection antibodies. Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) were employed to characterize the Cu and Cd nanocrystals. The covalently modified electrode with the Fab antibody fragments through the Au-thiolate bond (to dispel the non-specific adsorption) was investigated via scanning electron microscopy (SEM). After the sandwiched immunoreaction, the antibody-modified nanocrystals were captured on the immunosensor, which could be interrogated in pH 3.5 HCl using square-wave anodic stripping voltammetry. Experimental results also indicated that the multiplexed immunoassay enabled the simultaneous detection of HIgG and RIgG in a single run with the similar linear range from 0.01 to 10 ng mL -1 , and the limits of detection (LODs) towards two analytes could be as low as 3.4 pg mL -1 (at 3σ). Acceptable assay results on precision, reproducibility, specificity, and method accuracy were also acquired. Importantly, the newly designed strategy avoided cross-talk and enzymatic introduction as compared to conventional electrochemical immunoassays, thus exhibiting a promising potential in clinical applications.

Au nanoparticles (AuNPs) as signal reporters have been utilized in colorimetric in vitro diagnostics (IVDs) for decades. Nevertheless, it remains a grand challenge to substantially enhance the detection sensitivity of AuNP-based IVDs as confined by the inherent plasmonics of AuNPs. In this work, we circumvent this confinement by developing unique dual-functional AuNPs that were engineered by coating conventional AuNPs with ultrathin Pt skins of sub-10 atomic layers (i.e., Au@Pt NPs). The Au@Pt NPs retain the plasmonic activity of initial AuNPs while possessing ultrahigh catalytic activity enabled by Pt skins. Such dual functionalities, plasmonics and catalysis, offer two different detection alternatives: one produced just by the color from plasmonics (low-sensitivity mode) and the second more sensitive color catalyzed from chromogenic substrates (high-sensitivity mode), achieving an "on-demand" tuning of the detection performance. Using lateral flow assay as a model IVD platform and conventional AuNPs as a benchmark, we demonstrate that the Au@Pt NPs could enhance detection sensitivity by 2 orders of magnitude.

A non-enzyme cascade amplification strategy, based on the dissolution of Ag nanoparticles and a Pt nanocube-catalyzed reaction, for colorimetric assay of disease biomarkers was developed. This strategy overcomes the intrinsic limitations of enzymes involved in conventional enzymatic amplification techniques, thanks to the utilization of noble-metal nanostructures with superior properties.

Aflatoxin B 1 (AFB 1 ) monitoring has attracted extensive attention because food safety is a worldwide public health problem. Herein, we design a novel simultaneously visual and photoelectrochemical (PEC) immunosensing system for rapid sensitive detection of AFB 1 in foodstuff. The immunoreaction was carried out on anti-AFB 1 antibody-modified magnetic beads by using glucose oxidase (GOx)-labeled AFB 1 -bovine serum albumin (AFB 1 -BSA) conjugates as the tags with a competitive-type immunoassay format, while the visual and PEC evaluation was performed via carbon quantum dots (CQDs)-functionalized MnO 2 nanosheets. Accompanying the formation of immunocomplexes, the carried GOx initially oxidized the substrate (glucose) for the generation of H 2 O 2 , which reduced/etched MnO 2 nanosheets into Mn 2+ ions, thereby resulting in the dissociation of CQDs from the electrode. Within the applied potentials, the photocurrent of MnO 2 -CQDs-modified electrode decreased with the increasing H 2 O 2 level in the detection cell. Meanwhile, a visual detection could be performed according to the change in the color of MnO 2 -CQDs-coated electrode. To elaborate, this system was aggregated into a high-throughput microfluidic device to construct a semiautomatic detection cell. Under optimal conditions, the photocurrent increased with the increasing target AFB 1 within a dynamic working range from 0.01 to 20 ng mL -1 with a limit of detection (LOD) of 2.1 pg mL -1 (ppt). The developed immunoassay exhibited good reproducibility and acceptable accuracy. In addition, the method accuracy relative to AFB 1 ELISA kit was evaluated for analyzing naturally contaminated or spiked peanut samples, giving the well-matched results between two methods. Although our strategy was focused on the detection of target AFB 1 , it is easily extended to screen other small molecules or mycotoxins, thereby representing a versatile immunosensing scheme.

Methods based on nanostructures have been developed for potentiometric immunosensors, but most involve low sensitivity or weak signal output and are unsuitable for routine use in diagnosis. Herein, we devise an in-situ signal-amplification strategy for enhanced electrical readout of potentiometric immunosensor toward target prostate-specific antigen (PSA, one kind of cancer biomarkers), based on polyion oligonucleotide-labeled gold nanoparticles (AuNPs). To decrease the background signal, monoclonal anti-human PSA capture antibody was covalently conjugated onto an activated glassy carbon electrode via typical carbodiimide coupling. AuNPs heavily functionalized with the polyion oligonucleotides and polyclonal anti-PSA detection antibodies (pAb 2 -AuNP-DNA) were utilized as the signal-generation nanotags. In the presence of target PSA, a sandwich-type immunoreaction was executed between capture antibody and detection antibody on the electrode. The detectable signal derived from the shift in the electric potential as a result of the change in the surface charge before and after the antigen-antibody reaction. With target PSA increased, the captured pAb 2 -AuNP-DNA to the electrode accompanying detection antibody increased, thereby resulting in the change of the electrode potential. Due to numerous polyion oligonucleotides with the negative charge, the signal readout amplified. Under the optimal conditions, the shift in the output potential was proportional to the logarithm of target PSA concentration and displayed a dynamic linear range from 0.05 to 20 ng mL -1 with a detection limit of 13.6 pg mL -1 . An intermediate precision of ≤13.2% was accomplished with the batch-to-batch identification. The selectivity was acceptable. The method accuracy was evaluated for human serum specimens, and gave the consistent results between the potentiometric immunosensor and the referenced enzyme-linked immunosorbent assay (ELISA)., (Copyright © 2017 Elsevier B.V. All rights reserved.)

This work designs a new label-free aptasensor for the colorimetric determination of small molecules (adenosine 5'-triphosphate, ATP) by using visible gold nanoparticles as the signal-generation tags, based on target-triggered hybridization chain reaction (HCR) between two hairpin DNA probes. The assay is carried out referring to the change in the color/absorbance by salt-induced aggregation of gold nanoparticles after the interaction with hairpins, gold nanoparticles and ATP. To construct such an assay system, two hairpin DNA probes with a short single-stranded DNA at the sticky end are utilized for interaction with gold nanoparticles. In the absence of target ATP, the hairpin DNA probes can prevent gold nanoparticles from the salt-induced aggregation through the interaction of the single-stranded DNA at the sticky end with gold nanoparticles. Upon target ATP introduction, the aptamer-based hairpin probe is opened to expose a new sticky end for the strand-displacement reaction with another complementary hairpin, thus resulting in the decreasing single-stranded DNA because of the consumption of hairpins. In this case, gold nanoparticles are uncovered owing to the formation of double-stranded DNA, which causes their aggregation upon addition of the salt, thereby leading to the change in the red-to-blue color. Under the optimal conditions, the HCR-based colorimetric assay presents good visible color or absorbance responses for the determination of target ATP at a concentration as low as 1.0nM. Importantly, the methodology can be further extended to quantitatively or qualitatively monitor other small molecules or biotoxins by changing the sequence of the corresponding aptamer., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Developing simple, high-efficiency non-enzyme bioassays is of great importance for modern analytical systems, but remains a significant challenge. One promising route is to utilize highly efficient nanocatalysts with the exposure of active crystal facets. Herein, we for the first time propose a novel colorimetric immunoassay for the ultrasensitive detection of the human prostate-specific antigen (PSA) based on using a unique type of nanolabel - high-index {hk0} faceted platinum concave nanocubes (HIF-Pt-CNCs). The proposed HIF-Pt-CNCs exhibit superior peroxidase-like catalytic activity that is ∼1500- and ∼4-fold higher than that of natural horseradish peroxidase and Pt nanospheres, respectively, and thereby can provide powerful signal amplification by catalyzing the oxidation of peroxidase substrates in the presence of hydrogen peroxide. Using the HIF-Pt-CNC-labelled anti-PSA detection antibody as a signal probe, the immunoassay is carried out in anti-PSA capture antibody-immobilized microplate wells in a sandwich-type detection mode. Under optimal conditions, the developed immunoassay is able to achieve high sensitivity and specificity for PSA detection in a linear range of 20-2000 pg mL -1 and with an ultralow detection limit of 0.8 pg mL -1 , which is much lower than that of conventional enzyme-linked immunosorbent assay (ELISA). Moreover, the method is validated for the analysis of 10 PSA clinical serum specimens, and the results agree very well with those obtained by using a commercialized ELISA kit. Therefore, this new, facile and efficient immunoassay is a promising technique with potential applications in medical science research and clinical diagnosis.

Photoelectrochemical (PEC) measurement has been developed rapidly for bioanalysis in recent years. However, the actual application for most existed PEC bioanalytical systems is still a challenge because the perfect solutions for sensing surface design, high-throughput detection, and portability are lacked. To successfully overcome these limitations and realize accurate, continuous screening and assessing on prognostic indicator of early stage cancer on the spot, an innovative and portable semiautomated support power-free photoelectrochemical (SP-PEC) immunosensing platform consisted with a miniature semiautomatic injection system and digital multimeter (DMM) readout is designed (prostate specific antigen, PSA, was used as the proof-of-concept analyte). Decahedral BiVO 4 that decorated with Au nanocrystal on {010} facets (Au-BiVO 4 ) by photodeposition is used as the photoanode materials to produce photocurrent signal under irradiation of micro laser light (5.0 w, λ ≥ 380 nm). The monoclonal anti-PSA capture antibody (mAb 1 )-functionalized Fe 3 O 4 magnetic nanobeads (mAb 1 -MN) and glucose oxidase (GOx)/monoclonal detection antibody (mAb 2 )-conjugated gold nanoparticle (GOx-AuNP-mAb 2 ) are employed as immunosensing probe and signal probe, respectively. The H 2 O 2 as an excellent holes scavenger that in suit generated from GOx oxidization glucose substrate significantly amplifies the photocurrent. The variation of instantaneous current value that registered as the signal of the immunoassay increases linearly with the logarithm of target PSA concentration increasing in a wide range from 10 pg mL -1 to 100 ng mL -1 with a low detection limit (LOD) of 4.0 pg mL -1 . The SP-PEC immunosensing platform not only simplifies the assay process, but also improves detecting efficiency. The semiautomatic and portable SP-PEC analysis device allows analysis on spot and high-throughput continuous detection. Additional, we also gain deep insight into the relations between the specific shape as well as Au nanocrystal decoration and PEC activity and speculate the possible enhancement mechanisms of Au-BiVO 4 . Therefore, the present work not only develops a flexible SP-PEC biosensor platform for rapid and continuous detection, but also provides a possible route for designing high performance photoelectric materials.

This work reports on a novel time-resolved fluorescent aptasensing platform for the quantitative monitoring of adenosine 5'-triphosphate (ATP) by interaction of dispersive/agglomerate gold nanoparticles (AuNPs) with terbium ion-coordinated carbon dots (Tb-CDs). To construct such a fluorescent nanoprobe, Tb-CDs with high-efficient fluorescent intensity are first synthesized by the microwave method with terbium ions (Tb(3+)). The aptasensing system consists of ATP aptamer, AuNP and Tb-CD. The dispersive/agglomerate gold nanoparticles are acquired through the reaction of the aptamer with target ATP. Upon target ATP introduction, the aptamers bind with the analytes to form new aptamer-ATP complexes and coat on the surface of AuNPs to inhibit their aggregation in the high salt solution. In this case, the fluorescent signal of Tb-CDs is quenched by the dispersive AuNPs on the basis of the fluorescence resonance energy transfer (FRET). At the absence of target analyte, gold nanoparticles tend to aggregate in the high salt state even if the aptamers are present. Thus, the added Tb-CDs maintain their intrinsic fluorescent intensity. Experimental results indicated that the aptasensing system exhibited good fluorescent responses toward ATP in the dynamic range from 40nM to 4.0μM with a detection limit of 8.5nM at 3sblank criterion. The repeatability and intermediate precision is less than 9.5% at three concentrations including 0.04, 0.4 and 2.0μM ATP. The selectivity was acceptable toward guanosine 5'-triphosphate, uridine 5'-triphosphate and cytidine 5'-triphosphate. The methodology was applied to evaluate the blank human serum spiked with target ATP, and the recoveries (at 3 concentration levels) ranged between 97.0% and 103.7%. Importantly, this detection scheme is rapid, simple, cost-effective, and does not require extensive sample preparation or separation., (Copyright © 2016 Elsevier B.V. All rights reserved.)

In this work, a novel and sensitive sandwich-type electrochemical aptasensor has been developed for thrombin detection based on platinum nanoparticles (Pt NPs) decorated carbon nanocages (CNCs) as signal tags. The morphological and compositional of the Pt NPs/CNCs were examined using transmission electron microscopy, X-ray diffraction, and Raman spectroscopy. The results showed that the Pt NPs with about 3-5nm in diameter were well dispersed on the surface of CNCs. The thiolated aptamer was firstly immobilized on the gold electrode to capture the thrombin molecules, and then aptamer functionalized Pt NPs/CNCs nanocomposites were used to fabricate a sandwich sensing platform. Then, the high-content Pt NPs on carbon nanocages acting as hydrogen peroxide-mimicking enzyme catalyzed the reduction of H2O2, resulting in significant electrochemical signal amplification. Differential pulse voltammetry is employed to detect thrombin with different concentrations. Under optimized conditions, the approach provided a good linear response range from 0.05 pM to 20nM with a low detection limit of 10fM. This Pt NPs/CNCs-based aptasensor shows good precision, acceptable stability and reproducibility, which provided a promising strategy for electrochemical aptamer-based detection of other biomolecules., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Herein, gold-silver bimetallic nanoclusters (Au-Ag NCs) with the high fluorescent intensity were first synthesized successfully and utilized for the fabrication of sensitive and specific sensing probes toward inorganic pyrophosphatase (PPase) activity with the help of copper ion (Cu(2+)) and inorganic pyrophosphate ion (PPi). Cu(2+) was used as the quencher of fluorescent Au-Ag NC, while PPi was employed as the hydrolytic substrate of PPase. The system consisted of PPi, Cu(2+) ion, and bovine serum albumin (BSA)-stabilized Au-Ag NC. The detection was carried out by enzyme-induced hydrolysis of PPi to liberate copper ion from the Cu(2+)-PPi complex. In the absence of target PPase, free copper ions were initially chelated with inorganic pyrophosphate ions to form the Cu(2+)-PPi complexes via the coordination chemistry, thus preserving the natural fluorescent intensity of the Au-Ag NCs. Upon addition of target PPase into the detection system, the analyte hydrolyzed PPi into phosphate ions and released Cu(2+) ion from the Cu(2+)-PPi complex. The dissociated copper ions readily quenched the fluorescent signal of Au-Ag NCs, thereby resulting in the decrease of fluorescent intensity. Under optimal conditions, the detectable fluorescent intensity of the as-prepared Au-Ag NCs was linearly dependent on the activity of PPase within a dynamic linear range of 0.1-30 mU/mL and allowed the detection at a concentration as low as 0.03 mU/mL at the 3sblank criterion. Good reproducibility (CV < 8.5% for the intra-assay and interassay), high specificity, and long-term stability (90.1% of the initial signal after a storage period of 48 days) were also received by using our system toward target PPase activity. In addition, good results with the inhibition efficiency of sodium fluoride were obtained in the inhibitor screening research of pyrophosphatase. Importantly, this system based on highly enhanced fluorescent Au-Ag NCs offer promise for simple and cost-effective screening of target PPase activity without the needs of sample separation and multiple washing steps.

A new impedimetric sensing strategy based on enzyme-triggered formation of enzyme-tyramine concatamers on the nanogold-functionalized poly(amidoamine) (PAMAM) dendrimer was designed for sensitive detection of mercury(II) (Hg(2+)) ion, coupling with enzymatic biocatalytic precipitation towards 4-choloro-1-naphthol (4-CN) on thymine (T)-rich single-stranded DNA1-modified electrode. Initially, nanogold-decorated PAMAM dendrimer (AuNP-PAMAM) was synthesized by the in-situ reduction method, and then functionalized with horseradish peroxidase (HRP) and another T-rich oligomer (DNA2). Upon target Hg(2+) introduction, probe DNA2 on the AuNP-PAMAM bound to the DNA1 on the electrode owing to the T-Hg(2+)-T coordination chemistry between the two DNA strands. Accompanying the AuNP-PAMAM, the carried HRP could trigger the formation of HRP-tyramine concatamer via the classical tyramine signal amplification strategy in the presence of HRP-tyramine conjugates and hydrogen peroxide. The concatenated HRP molecules in the concatamer catalyzed the 4-CN oxidation to produce an insoluble precipitation on the electrode, thereby resulting in the local alteration in the conductivity. Under optimal conditions, two signal-generation tags including HRP-AuNP-DNA2 and HRP-AuNP-PAMAM-DNA2 with or without tyramine signal amplification strategy (i.e., four schemes) were used for impedimetric detection of target Hg(2+) on the basis of the same assay format. A low detection limit (LOD) of 0.4pM and a wide dynamic working range of 0.001-100nM Hg(2+) by using HRP-AuNP-PAMAM-DNA2 with tyramine signal amplification strategy were obtained in comparison with those of other strategies. The assay had a good repeatability and showed an intermediate precision of down to 9.6%. In addition, the methodology also exhibited high specificity and selectivity towards target Hg(2+) against other metal ions, and was applicable for monitoring Hg(2+) in the spiked drinking water samples., (Copyright © 2015 Elsevier B.V. All rights reserved.)

How to concentrate target molecules on the surface of a SERS substrate is a key problem in the practical application of SERS. Herein, we designed for the first time a pollutant capturing surface enhanced Raman spectroscopy (SERS) substrate, namely porous BN microfibers uniformly decorated with Ag nanoparticles, in which the BN microfibers adsorb pollutants, while the Ag nanoparticles provide SERS activity. This SERS substrate captures pollutants from an aqueous solution completely and accumulates them all on its surface without introducing noise signals. The pores of BN protect the silver particles from aggregation which makes BN/Ag a stable and recyclable SERS substrate. What's more, while the dyes are thoroughly concentrated from a diluted solution, the SERS detection limit is easily enhanced, from 10(-6) M to 10(-9) M.

5,10,15,20-Tetraphenyl-21H,23H-porphine cobalt flat stacking on the reduced graphene oxide with platinum nanoparticles (PtNPs/CoTPP/rGO) were first synthesized and functionalized with monoclonal rabbit anti-aflatoxin B1 antibody (anti-AFB1) for highly efficient electrochemical immunoassay of aflatoxin B1 (AFB1) in this work. Transmission electron microscopy (TEM), atomic force microscope (AFM) and spectral techniques were employed to characterize the PtNPs/CoTPP/rGO hybrids. Using anti-AFB1-conjugated PtNPs/CoTPP/rGO as the signal-transduction tag, a novel non-enzymatic electrochemical immunosensing system was designed for detection of target AFB1 on the AFB1-bovine serum albumin-functionalized sensing interface. Experimental results revealed that the designed immunoassay could exhibit good electrochemical responses for target analyte and allowed the detection of AFB1 at a concentration as low as 5.0 pg mL(-1) (5.0 ppt). Intra- and inter-assay coefficients of variation were below 10%. Importantly, the methodology was further validated for analyzing naturally contaminated or spiked blank peanut samples with consistent results obtained by AFB1 ELISA kit, thus providing a promising approach for quantitative monitoring of organic pollutants.

The development of signal-amplified colorimetric immunoassay relies on the design of highly efficient signal-transduction tags. One promising route is to exploit a novel enzyme mimetic system as the signal label. Herein, we report that urchin-like (gold core)@(platinum shell) nanohybrids (Au@PtNHs) can be utilized as a highly efficient peroxidase mimetic system for in situ amplified colorimetric immunoassay of prostate-specific antigen (PSA, one kind of tumor marker). Initially, urchin-like Au@PtNHs were discovered to outperform horseradish peroxidase (HRP) by a vast margin in terms of the turnover number toward hydrogen peroxide (H2O2)-3,3',5,5'-tetramethylbenzidine (TMB) system and the stability against high temperatures and HRP inhibitors. Based on this discovery, the assay was simply carried out on a capture antibody-immobilized microplate by using the Au@PtNH-labeled detection antibody as a signal-transduction tag with a sandwich-type assay mode. The colorimetric signal stemmed from the labeled Au@PtNHs toward catalytic oxidation of TMB-H2O2 system. Experimental results indicated that the Au@PtNH-based colorimetric immunoassay could display a good colorimetric response toward PSA in the dynamic working range of 5-500 pg mL(-1) with a low detection limit of 2.9 pg mL(-1). Meanwhile, the developed immunoassay exhibited good precision and reproducibility, high specificity and acceptable accuracy for the detection of clinical serum samples. These results open up a new horizon for the development of highly sensitive, highly stable and inexpensive non-enzyme immunoassay platforms as an alternative to conventional enzyme-based immunoassay platforms., (Copyright © 2015 Elsevier B.V. All rights reserved.)