High-Performance Detection of Exosomes Based on Synergistic Amplification of Amino-Functionalized Fe3O4 Nanoparticles and Two-Dimensional MXene Nanosheets.

Exosomes derived from cancer cells have been recognized as a promising biomarker for minimally invasive liquid biopsy. Herein, a novel sandwich-type biosensor was fabricated for highly sensitive detection of exosomes. Amino-functionalized Fe3O4 nanoparticles were synthesized as a sensing interface with a large surface area and rapid enrichment capacity, while two-dimensional MXene nanosheets were used as signal amplifiers with excellent electrical properties. Specifically, CD63 aptamer attached Fe3O4 nanoprobes capture the target exosomes. MXene nanosheets modified with epithelial cell adhesion molecule (EpCAM) aptamer were tethered on the electrode surface to enhance the quantification of exosomes captured with the detection of remaining protein sites. With such a design, the proposed biosensor showed a wide linear range from 102 particles µL-1 to 107 particles µL-1 for sensing 4T1 exosomes, with a low detection limit of 43 particles µL-1. In addition, this sensing platform can determine four different tumor cell types (4T1, Hela, HepG2, and A549) using surface proteins corresponding to aptamers 1 and 2 (CD63 and EpCAM) and showcases good specificity in serum samples. These preliminary results demonstrate the feasibility of establishing a sensitive, accurate, and inexpensive electrochemical sensor for detecting exosome concentrations and species. Moreover, they provide a significant reference for exosome applications in clinical settings, such as liquid biopsy and early cancer diagnosis.

Facile bimetallic co-amplified electrochemical sensor for folic acid sensing based on CoNPs and CuNPs.

Folic acid (FA) is essential for human health, particularly for pregnant women and infants. In this work, a glassy carbon electrode (GCE) was modified by a bimetallic layer of Cu/Co nanoparticles (CuNPs/CoNPs) as a synergistic amplification element by simple step-by-step electrodeposition, and was used for sensitive detection of FA. The proposed CuNPs/CoNPs/GCE sensor was characterized by differential pulse voltammetry (DPV), electrochemical impedance spectroscopy (EIS) and field emission scanning electron microscopy (FESEM). Then, under optimal conditions, a linear relationship was obtained in the wide range of 110.00-1750.00 µM for the detection of FA with a limit of detection (LOD) of 34.79 µM (S/N = 3). The sensitivity was calculated as 0.096 µA µM-1 cm-2. Some interfering compounds including glucose (Glc), biotin, dopamine (DA), and glutamic acid (Glu) showed little effect on the detection of FA by amperometry (i-t). Finally, the average recovery obtained was in a range of 91.77-110.06%, with a relative standard deviation (RSD) less than 8.00% in FA tablets, indicating that the proposed sensor can accurately and effectively detect the FA content in FA tablets.

Sandwich-type microRNA biosensor based on graphene oxide incorporated 3D-flower-like MoS2 and AuNPs coupling with HRP enzyme signal amplification.

A sandwich electrochemical biosensing strategy for ultrasensitive detection of miRNA-21 was developed by using graphene oxide incorporated 3D-flower-like MoS2 (3D MoS2-rGO) nanocomposites as the substrate and horseradish peroxidase (HRP)-functionalized DNA strand 1 (S1)-gold nanoparticles (S1-AuNPs-HRP) as signal amplification probes. Herein, 3D MoS2-rGO nanocomposites not only had a large specific surface area and excellent conductivity, but also provided more attachment sites for electrodepositing AuNPs. In the presence of target miRNA, a sandwich structure was formed, and the determination of the miRNA-21 was carried out by measuring the DPV response of H2O2 mediated by hydroquinone (HQ) at a potential of + 0.052 V (vs AgCl reference electrode). Under the optimal experimental conditions, the as-prepared biosensor enabled the ultrasensitive detection of miRNA-21 from 5 fM to 0.5 µM with the low detection limit of 0.54 fM (S/N = 3), comparable or lower than previous reported methods for miRNA-21 detection, which benefited from the synergistic amplification of 3D MoS2-rGO and AuNPs-HRP. The prepared biosensor showed satisfactory selectivity, reproducibility, and stability towards miRNA-21 detection. The biosensor was feasible for accurate and quantitative detection of miRNA-21 in normal human serum samples with RSD below 5.86%, which showed a great potential in clinical analysis and disease diagnosis.

Ps-Pt nanozyme-based synergistic signal amplification biosensor for highly sensitive colorimetric detection of protein.

Immunosorbent assay is one of the most popular immunological screening techniques which has been widely used for the clinical diagnosis of alpha-fetoprotein (AFP). While traditional immunosorbent assay (ELISA) suffers from low detection sensitivity due to its low intensity of colorimetric signal. To improve the sensitivity of AFP detection, we developed a new and sensitive immunocolorimetric biosensor by combining Ps-Pt nanozyme with terminal deoxynucleotidyl transferase (TdT)-mediated polymerization reaction. The determination of AFP was achieved by measuring the visual color intensity produced by the catalytic oxidation reaction of the 3,3',5,5'-tetramethylbenzidine (TMB) solution with Ps-Pt and horseradish peroxidase (HRP). Owing to the synergistic catalysis of Ps-Pt and horseradish peroxidase HRP enriched in polymerized amplification products, this biosensor exhibited a significant color change within 25 s in the presence of 10-500 pg/mL AFP. This proposed method allowed for the specific detection of AFP with a detection limit of 4.30 pg/mL and even 10 pg/mL target protein could be distinguished clearly by visual observation. Furthermore, this biosensor could be applied to analysis of AFP in the complex sample and could be easily extended to the detection of other proteins.

Hollow SnO2/CdS QDs/CdCO3 heterostructured nanocubes coupled with hollow PtPd/MnCo-CeO2 nanozyme-mediated synergistic amplification for ultrasensitive PEC immunoanalysis of lung cancer biomarker.

Nowadays, lung cancer is one of the most dangerous cancers threatening human life all over the world. As a crucial biomarker, cytokeratin 19 fragment 21-1 (CYFRA 21-1) is extraordinary important for diagnosis of non-small cell lung cancer (NSCLC). In this work, we synthesized hollow SnO2/CdS QDs/CdCO3 heterostructured nanocubes with high and stable photocurrents, which applied to construction of a sandwich-typed photoelectrochemical (PEC) immunosensor for detection of CYFRA 21-1, integrated by in-situ catalytic precipitation strategy with home-built PtPd alloy anchored MnCo-CeO2 (PtPd/MnCo-CeO2) nanozyme for synergistic amplification. The interfacial electron transfer mechanism upon visible-light irradiation was investigated in details. Further, the PEC responses were seriously quenched by the specific immunoreaction and precipitation catalyzed by the PtPd/MnCo-CeO2 nanozyme. The established biosensor showed a wider linear range of 0.001-200 ng mL-1 and a lower limit of detection (LOD = 0.2 pg mL-1, S/N = 3), coupled by exploring such analysis even in diluted human serum sample. This work opens a constructive avenue to develop ultrasensitive PEC sensing platforms for detecting diverse cancer biomarkers in clinic.

Cu-MOFs/GOx Bifunctional Probe-Based Synergistic Signal Amplification Strategy: Toward Highly Sensitive Closed Bipolar Electrochemiluminescence Immunoassay.

A closed bipolar electrochemiluminescence (BP-ECL) platform for sensitive prostate specific antigen (PSA) detection was proposed based on a novel synergistic signal amplification strategy. Specifically, glucose oxidase-loaded Cu-based metal-organic frameworks (Cu-MOFs/GOx) as bifunctional probes were bridged on the anodic interface with the target PSA as the intermediate unit. In virtue of the large loading capacity of Cu-MOFs, a large amount of a co-reactant, i.e., H2O2 in this L-012-based ECL system and gluconic acid were generated on the anodic pole in the presence of glucose. The generated gluconic acid could effectively degrade the Cu-MOFs to release Cu2+ which greatly accelerates the formation of highly active intermediates from co-reactant H2O2, boosting the ECL intensity. As for the cathodic pole, K3Fe(CN)6 with a lower reduction potential is used to reduce the driving voltage and speed up the reaction rate, further strengthening the ECL intensity. Thanks to the synergistic signal amplification effect at both two electrode poles of the BP-ECL system, highly sensitive detection of PSA was realized with a detection limit of 5.0 × 10-14 g/mL and a wide linear range of 1.0 × 10-13-1.0 × 10-7 g/mL. The strategy provides a novel way for signal amplification in the BP-ECL biosensing field.

Picomolar thrombin detection by orchestration of triple signal amplification strategy with hierarchically porous Ti3C2Tx MXene electrode material-catalytic hairpin assembly reaction-metallic nanoprobes.

Signal amplification strategies are essential to boost the sensitivity of detecting targeted ions or molecules with important biological functions, while few studies take advantage of signal amplification strategies more than two. As a "proof-of-concept" demonstration, we present the ultrasensitive electrochemical aptasensor for picomolar thrombin detection by synchronous coordination of triple signal amplification strategy. The porous MXene framework (PMXF) with secondary pores is constructed as carrier to increase electrons transfer channels, and thionine (as redox indicator) labelled Au nanorod (AuNR) and hollow Cu-Pt alloy (HCuPtA) are synthesized as the electrical signal amplifiers to enhance the response signals. In the presence of picomolar-level thrombin, catalytic hairpin assembly reactions of DNA are triggered to bridge thionine labelled AuNR or HCuPtA nanoprobes on the PMXF with controllablly scondary pore structures. Under the optimal conditionals, the sandwich-typed aptasensor based on PMXF-5/AuNR shows a more low limit of detection (LOD) of 0.67 pM with a linear range from 2 pM to 10 nM, while PMXF-5/HcuPtA exhibits a more wide linear range from 50 pM to 50 nM with a LOD of 16.67 pM for thormbin. This sensing platform can be customized to analyze other biological or environmental substances at an ultrahigh level by rationally designing DNA sequences of target-binding aptamer.

High-performance electrochemical mercury aptasensor based on synergistic amplification of Pt nanotube arrays and Fe3O4/rGO nanoprobes.

In this work, a novel sandwich-type aptasensor was designed for the ultrasensitive recognition of trace mercury ions in water. Numerous oriented platinum nanotube arrays (PtNAs) were in-situ crystallized on a flexible electrode as a sensing interface, while thionine labelled Fe3O4/rGO nanocomposites as signal amplifiers. Both PtNAs/CF and nanocomposites were synthesized by easy hydrothermal processes. With their large surface area, it was favorable for electrochemical performance and immobilization of capture DNAs (cDNA) and report DNAs (rDNA). Upon the existence of Hg2+, partial linker DNAs were tightly bound with cDNAs through thymine-Hg2+-thymine pairing (T-Hg2+-T). Then rDNAs attached Fe3O4/rGO nanoprobes were fixed on the electrode through the match of remaining linker DNAs and rDNAs. Under the optimal conditions, the Hg2+ aptasensor showed a synergistic amplification performance with a wide linear range from 0.1nM to 100nM, as well as a low detection limit of 30pM. Moreover, the as-prepared aptasensor also exhibited reliable performance for assay in real lake water samples.