A highly sensitive dual-mode detection platform based on the novel copper/molybdenum bimetallic nanoclusters and Co-Fe layered doubled hydroxide nanozyme for butyrylcholinesterase activity sensing.

Herein, a novel copper/molybdenum bimetallic nanoclusters (Cu/Mo NCs) with intense blue emission were synthesized by using polyvinylpyrrolidone (PVP) as template and ascorbic acid as reducing agent. Owing to the synergistic effect between Cu and Mo, the fluorescence intensity of Cu/Mo NCs was significantly improved about 6-time than monometallic copper nanoclusters. A novel and sensitive ratiometric fluorescence and colorimetric dual-mode sensing platform for monitoring butyrylcholinesterase (BChE) was strategically constructed by the integration of Cu/Mo NCs with excellent optical properties and Co-Fe layered doubled hydroxide (CoFe-LDH) with superior peroxidase-like activity for the first time. In the presence of H2O2, nonfluorescent and colorless o-phenylenediamine (OPD) was oxidized to fluorescent and yellow 2,3-diaminophenazine (DAP) with maximum fluorescence emission peak at 564 nm and ultraviolet absorption peak at 418 nm by CoFe-LDH with peroxidase-like activity. Simultaneously, the generation of DAP could effectively quench Cu/Mo NCs fluorescence at 444 nm through the inner-filter effect (IFE). The hydrolysis of S-butyrylthiocholine iodide (BTCh) can be catalyzed by butyrylcholinesterase (BChE) to generate thiocholine (TCh) that could hinder the oxidation of OPD, leading to the fluorescence and ultraviolet absorption of DAP decreased, meanwhile, the fluorescence of Cu/Mo NCs recovered. The ratiometric fluorescence signal F564/F444 and colorimetric system both performed a satisfactory response to the concentration of BChE in the range 0.5 to 90 U L-1 and 1 to 100 U L-1 with the LOD of 0.18 U L-1 and 0.36 U L-1, respectively. The dual-mode sensing for BChE exhibited outstanding application potential in biosensing.

High-performance colorimetric sensor based on PtRu bimetallic nanozyme for xanthine analysis.

The identification and quantification of xanthine are crucial for assessing the freshness and quality of food products, particularly in the seafood industry. Herein, a new approach was developed, involving the in-situ controllable growth of Pt91Ru9 nanoparticles on graphitic carbon nitride to yield Pt91Ru9@C3N4 catalytic materials. By integrating Pt91Ru9@C3N4 with the xanthine/xanthine oxidase (XOD) enzyme catalytic system, a nanozyme-enzyme tandem platform was obtained for the quantification analysis of xanthine. Under the catalytic oxidation of xanthine by XOD in the presence O2, H2O2 was generated. Upon the addition of peroxidase-like activity of Pt91Ru9@C3N4, H2O2 can be decomposed into •OH and 1O2, which can further catalyze the oxidation of TMB to its oxidation product oxTMB with an absorption peak at 652 nm. This smartphone-assisted portable colorimetric sensor for visual monitoring xanthine with a low detection limit of 8.92 nmol L-1, and successfully applied to detect xanthine in grass carp and serum samples.

Construction of the fluorescence sensing platform with a bifunctional Cu@MOF nanozyme for determination of alkaline phosphatase and its inhibitor.

In this work, a novel and sensitive fluorescence sensing system for alkaline phosphatase (ALP) was constructed using a bifunctional copper metal-organic framework (Cu@MOF) nanozyme, which had excellent oxidase-mimetic activity and fluorescence properties. Owing to the presence of 2-amino-1,4-benzenedicarboxylic acid (1,4-BDC-NH2) ligand, Cu@MOF displays excellent fluorescence performance at 444 nm. Additionally, Cu2+ endows the oxidase-like activity of Cu@MOF, which could trigger p-phenylenediamine (PPD) to be oxidized to a brown product (PPDox) and quench the photoluminescence of Cu@MOF through the inner filtration effect (IFE). As the preferential affinity of ATP for Cu2+, the catalytic activity of Cu@MOF was significantly reduced once ATP was added, thus PPD could not be oxidized and fluorescence was recovered. In the presence of ALP, ATP was hydrolyzed to adenosine and Pi, which allowed Cu@MOF to regain its catalytic activity and continued to catalyze the generation of PPDox. The fluorescence of Cu@MOF was therefore weakened once again. The ALP activity was directly proportional to the degree of decrease in fluorescence intensity. Thus, this novel fluorescence sensing strategy had a linear range of 0.5-60 U/L and the limit of detection was 0.14 U/L. The established sensing method could also be used to for ALP inhibitors screening, and achieved satisfactory results in determining the level of ALP activity in human serum.

A novel robust hydrogel-assisted paper-based sensor based on fluorescence UiO-66-NH2@ZIF-8 for the dual-channel detection of captopril.

Captopril (CP) is commonly used as an active enzyme inhibitor for the treatment of coronary heart disease, hypertension and angina pectoris. The development of sensitive and efficient method for CP analysis is of great importance in biomedical research. Herein, we fabricated a sensitive and robust hydrogel-assisted paper-based sensor based on fluorescence UiO-66-NH2@ZIF-8 and Co, N-doped carbon nanozymes with oxidase-mimicking activity for accurate monitoring of captopril. The hydrogel-assisted paper-based sensor appeared a visible pink signal due to the catalytic oxidation of colorless N,N-diethyl-p-phenylenediamine (DPD) to oxDPD by Co, N-doped carbon-based nanozymes, and resulted in the fluorescence quenching of UiO-66-NH2@ZIF-8. In the presence of captopril, the oxidation of chromogenic substrate DPD by Co, N-doped nanozymes in the hydrogel-assisted paper-based sensor was hindered and accompanied by a change in the visible color, leading to recovery of the fluorescence of UiO-66-NH2@ZIF-8, and the change in the fluorescence color could also be observed. Therefore, the quantitative detection of captopril is achieved by taking a smartphone photograph and converting the image parameters into data information using ImageJ software. The portable hydrogel-assisted paper sensor provided sensitive detection of captopril in two modes based on visible color change as well as fluorescence color change with limits of detection of 0.45 µM and 0.47 µM, respectively. This hydrogel-assisted paper-based sensor has been successfully applied to the accurate monitoring of captopril in human serum, providing a potential avenue for in situ detection of captopril.

Fabrication of superior laccase-mimicking enzyme with catalytic oxidative and photothermal properties for anti-bacterial and dual-mode glutathione S-transferase monitoring.

A novel laccase mimic enzyme Cu-Mn with excellent photothermal properties was firstly prepared via a combination of hydrothermal and in situ synthesis. Cu-Mn nanozymes could catalyze the typical laccase substrate 2,4-dichlorophenol (2,4-DP) to generate the red quinone imine. Further, loading the MnO2 nanosheets with photothermal properties, Cu-Mn nanozymes possessed not only excellent laccase catalytic activity, but also high photothermal conversion efficiency. The presence of glutathione S-transferase (GST) recovered the glutathione (GSH)-induced weakness of the laccase activity and photothermal properties of Cu-Mn. Hence, a GST enzyme-regulated dual-mode sensing strategy was established based on Cu-Mn nanozymes. The detection limits of GST monitoring based on colorimetric and photothermal methods were 0.092 and 0.087 U/L with response times of 20 min and 8 min, respectively. Furthermore, the proposed method enabled the measuring of GST levels in human serum and was successfully employed in the primary evaluation of hepatitis patients. Another attraction, the impressive photothermal behavior also endowed the Cu-Mn nanozymes with promising antimicrobial properties, which exhibited significant antimicrobial effects against Escherichia coli (E.coli) and Staphylococcus aureus (S.aureus). Unsurprisingly, multifunctional Cu-Mn nanozymes certainly explore new paths in biochemical analysis and antimicrobial applications.

Boron-doped g-C3N4 supporting Cu nanozyme for colorimetric-fluorescent-smartphone detection of α-glucosidase.

BACKGROUND: Due to that the higher activity of nanozymes would bring outstanding performance for the nanozyme-based biosensing strategies, great efforts have been made by researchers to improve the catalytic activity of nanozymes, and novel nanozymes with high catalytic activity are desired. Considering the crucial role in controlling blood glucose level, strategies like colorimetric and chemiluminescence to monitor α-glucosidase are developed. However, multi-mode detection with higher sensitivity was insufficient. Therefore, developing triple-mode detection method for α-glucosidase based on great performance nanozyme is of great importance. RESULTS: In this work, a novel nanozyme Cu-BCN was synthesized by loading Cu on boron doped carbon substrate g-C3N4 and applied to the colorimetric-fluorescent-smartphone triple-mode detection of α-glucosidase. In the presence of H2O2, Cu-BCN catalyzed the generation of 1O2 from H2O2, 1O2 subsequently oxidized TMB to blue colored oxTMB. In the presence of hydroquinone (HQ), the ROS produced from H2O2 was consumed, inhibiting the oxidation of TMB, which endows the possibility of colorimetric and visual on-site detection of HQ. Further, due to that the fluorescence of Mg-CQDs at 444 nm could be quenched by oxTMB, HQ could also be quantified through fluorescent mode. Since α-glucosidase could efficiently hydrolyze α-arbutin into HQ, the sensitive detection of α-glucosidase was realized. Further, colorimetric paper-based device (c-PAD) was fabricated for on-site α-glucosidase detection. The LODs for α-glucosidase via three modes were 2.20, 1.62 and 2.83 U/L respectively, high sensitivities were realized. SIGNIFICANCE: The nanozyme Cu-BCN possesses higher peroxidase-like activity by doping boron to the substrate than non-doped Cu-CN. The proposed triple-mode detection of α-glucosidase is more sensitive than most previous reports, and is reliable when applied to practical sample. Further, the smartphone-based colorimetric paper-based analytical device (c-PAD) made of simple materials could also detect α-glucosidase sensitively. The smartphone-based on-site detection provided a convenient, instrument-free and sensitive sensing method for α-glucosidase.

Construction of a dual-signal readout platform for effective glutathione S-transferase sensing based on polyethyleneimine-capped silver nanoclusters and cobalt-manganese oxide nanosheets with oxidase-mimicking activity.

A novel dual-mode fluorometric and colorimetric sensing platform is reported for determining glutathione S-transferase (GST) by utilizing polyethyleneimine-capped silver nanoclusters (PEI-AgNCs) and cobalt-manganese oxide nanosheets (CoMn-ONSs) with oxidase-like activity. Abundant active oxygen species (O2•-) can be produced through the CoMn-ONSs interacting with dissolved oxygen. Afterward, the pink oxDPD was generated through the oxidation of colorless N,N-diethyl-p-phenylenediamine (DPD) by O2•-, and two absorption peaks at 510 and 551 nm could be observed. Simultaneously, oxDPD could quench the fluorescence of PEI-AgNCs at 504 nm via the inner filter effect (IFE). However, in the presence of glutathione (GSH), GSH prevents the oxidation of DPD due to the reducibility of GSH, leading to the absorbance decrease at 510 and 551 nm. Furthermore, the fluorescence at 504 nm was restored due to the quenching effect of oxDPD on decreased PEI-AgNCs. Under the catalysis of GST, GSH and1-chloro-2,4-dinitrobenzo (CDNB) conjugate to generate an adduct, initiating the occurrence of the oxidation of the chromogenic substrate DPD, thereby inducing a distinct colorimetric response again and the significant quenching of PEI-AgNCs. The detection limits for GST determination were 0.04 and 0.21 U/L for fluorometric and colorimetric modes, respectively. The sensing platform illustrated reliable applicability in detecting GST in real samples.

A two-dimensional iron-doped carbon-based nanoenzyme with catalase-like activity for the detection of alkaline phosphatase and ascorbate oxidase.

Herein, we successfully synthesized two-dimensional iron-doped carbon-based nanosheets (Fe-N800 CS) with catalase-like activity through doping Fe into Zn MOF and introducing graphitic C3N4 (g-C3N4). The interaction of the Fe-N800 CS with hydrogen peroxide could generated abundant reactive oxygen species (ROS) and further oxidize o-Phenylenediamine (OPD) to 2,3-diaminophenazine (DAP) which has constant fluorescence at 560 nm. Ascorbic acid (AA) could be generated via the hydrolysis reaction between alkaline phosphatase (ALP) and ascorbic acid 2-phosphate (AAP). AA can be oxidized to dehy-droascorbic acid (DHA) by ROS, and then combined with OPD to generate 3-(1,2-dihydroxyethyl)furo[3,4b]-quinoxaline (QXD) with fluorescence at 440 nm, which could increase as the concentration of AA enhanced. DHA could also be generated through oxidation of AA by ascorbate oxidase (AAO). Thus, by monitoring the fluorescence ratio (I560/I440), a ratiometric fluorescence biosensing platform for ALP and AAO was established with the linear ranges in 0.2-10 U/L and 1-60 U/L, respectively. The limit of detection for ALP and AAO were 0.12 U/L and 0.59 U/L. Furthermore, the biosensing platform was successfully applied for the detection of ALP and AAO activity in human serum samples. This work provides a potential tool for future biomedical diagnostics.

A novel self-assembled dual-emissive ratiometric fluorescent nanoprobe for alkaline phosphatase sensing.

BACKGROUND: Alkaline phosphatase (ALP) is widely found in various organs and tissues of the human body which could assist in the verification of the presence of various diseases through its content in the blood. In the past few years, many analytical methods for ALP activity assays have been explored. However, a simple and economical method with high sensitivity and specificity also remains great challenge. Therefore, the development of sensitive and efficient approach for ALP analysis is of great significance in biomedical studies. RESULTS: Herein, we constructed a highly sensitive and label-free ratiometric fluorometric biosensing platform for the determination of ALP activity, which utilizing lysozyme(Ly)-functionalized 5-methyl-2-thiouracil(MTU)-modified gold nanoclusters (MTU-Ly@Au NC) and poly-dopamine (PDA) as signal indicators. Dopamine (DA) can self-polymerizes to form PDA under alkaline conditions that can further quenched the fluorescence of MTU-Ly@Au NC at 525 nm due to fluorescence resonance energy transfer (FRET) and absorption competition quenching (ACQ) effects. In this process, the PDA fluorescence intensity at 325 nm was nearly unchanged. After the addition of ALP, ascorbic acid (AA) which can alleviate the self-polymerization process of DA was generated from the substrate ascorbic acid 2-phosphate (AAP), thus changing ratiometric fluorescence intensity of I525/I325. Hence, by monitoring the fluorescence ratio (I525/I325), a ratiometric fluorescence biosensing platform for ALP was established with the linear calibration in the range of 0.5-8 U L-1 and the limit of detection of 0.157 U L-1. SIGNIFICANCE: This work not only synthesized a novel fluorescence probe with simple preparation and low cost for ALP which has excellent anti-interference properties and selectivity. Furthermore, this biosensing platform was successfully applied for the determination of ALP activity in human serum samples. This work provided a potential tool for biomedical diagnostics in the future.

Hydrogel-involved portable colorimetric sensor based on oxidase mimic Fe/Co-NC for acetylcholinesterase detection and pesticides inhibition assessment.

Herein, we synthesized a novel N-doped carbon layer encapsulated Fe/Co bimetallic nanoparticles (Fe/Co-NC), which exhibited superior oxidase-like activity due to the facilitation of electron penetration and the formation of metal-nitrogen active sites. Fe/Co-NC could catalyze the oxidation of 3,3,5,5-tetramethylbenzidine (TMB) to blue oxTMB. Acetylcholinesterase (AChE) could catalyze the hydrolysis of thioacetylcholine to produce reducing thiocholine, which prevented TMB from oxidation. Thus, a portable hydrogel colorimetric sensor was developed for on-site and visual monitoring of AChE with the detection limit of 0.36 U L-1, and successfully applied to detect AChE in human erythrocyte samples. Furthermore, this platform was used to investigate the inhibition of triazophos on AChE activity.

Self-Assembled Integrated Nanozyme Cascade Biosensor with Dual Catalytic Activity for Portable Urease Analysis.

In this work, a novel nanozyme (Cu@Zr) with all-in-one dual enzyme and fluorescence properties is designed by simple self-assembly. A nanozyme cascade sensor with disodium phenyl phosphate (PPDS) as substrate was first established by exploiting the dual enzymatic activities of phosphatase and laccase. Specifically, phosphatase cleaves the P-O bond of PPDS to produce colorless phenol, which is then oxidized by laccase and complexed with the chromogenic agent 4-aminoantipyrine (4-AP) to produce red quinoneimine (QI). Strikingly, the NH3 produced by the urease hydrolysis of urea can interact with Cu@Zr, accelerating the electron transfer rate and ultimately leading to a significantly improved performance of the cascade reaction. Moreover, the fluorescence at 440 nm of Cu@Zr is further quenched by the inner filter effect (IFE) of QI. Thus, the colorimetric and fluorescence dual-mode strategy for sensitive urease analysis with LODs of 3.56 and 1.83 U/L was established by the proposed cascade sensor. Notably, a portable swab loaded with Cu@Zr was also prepared for in situ urease detection with the aid of a smartphone RGB readout. It also provides a potentially viable analytical avenue for environmental and biological analysis.

Multi-signal aptasensor for thrombin detection based on catalytically active gold nanoparticles and fluorescent silicon quantum dots.

A multi-signal aptasensor for thrombin determination is proposed based on catalytically active gold nanoparticles (AuNPs) and fluorescent silicon quantum dots (SiQDs). Yellow 4-Nitrophenol (4-NP) could be converted to colorless 4-Aminophenol (4-AP) by catalytically active aptamer-modified AuNPs (S1-AuNPs). The SiQDs emitted strong blue fluorescence at 455 nm at the excitation wavelength of 367 nm. When thrombin was absent, S1-AuNPs could catalytically reduce yellow 4-NP to colorless 4-AP. When thrombin was added, the aptamer could be transformed into a G-quadruplex structure, which masked the surface-active catalytic sites of AuNPs and restrained the reduction of 4-NP. Thus, the fluorescence of SiQDs was greatly quenched by 4-NP through the inner filter effect (IFE), and the solution color remained yellow. As the concentration of thrombin increased, the catalytic activity of S1-AuNPs decreased. The concentration of 4-NP that was converted to 4-AP declined and the unconverted 4-NP increased. In this process, the absorption peak of 4-NP at 400 nm increased while the fluorescence emission of SiQDs at 455 nm decreased. The linear ranges of the fluorometric and colorimetric aptasensor were 0.5-30 nM and 0.3-30 nM, respectively. The limits of detection (LOD) for the two modes were 0.15 nM and 0.13 nM. Furthermore, a portable sensing platform was constructed by combining the smartphone-based device with the software ImageJ for the determination of thrombin. With the advantages of cost-effectiveness, simplicity of operation and broad applicability, this aptasensor provided a new perspective for on-site determination of thrombin in the clinical field.

A highly sensitive fluorescence "on-off-on" sensing platform for captopril detection based on AuNCs@ZIF-8 nanocomposite.

Here, a novel fluorescent sensing strategy is established for the detection of captopril (CP) sensitively on the basis of a nanocomposite of gold nanoclusters (AuNCs) and metal-organic framework (AuNCs@ZIF-8). The aggregation-induced emission (AIE) effect will be triggered when AuNCs is encapsulated by metal-organic framework (MOF) which served as a carrier since it limits the molecular motion of AuNCs, and the fluorescence of AuNCs greatly enhanced about 5-time after forming the nanocomposites of AuNCs@ZIF-8. The strong orange-emission at 562 nm was quenched in the presence of mercury ions through dynamic quenching. After adding captopril, the quenched fluorescence of AuNCs@ZIF-8/Hg2+ system would be restored due to the specific interaction among captopril with mercury ions. Simultaneously, the restored degree of AuNCs@ZIF-8/Hg2+ fluorescence depended on the concentration of captopril. Hence, with AuNCs@ZIF-8 serving as reporter signal, the captopril content can be monitored by an "on-off-on" fluorescence sensing mode with a linear relationship of 1-100 µM, and the limit of detection for captopril was 0.134 µM.

Construction of a Label-Free Ratiometric Biosensor Based on Target Recycling Amplification and Hg-ZnSe QDs for Assay of BChE and OPs.

Herein, we constructed a label-free ratiometric fluorescence biosensing strategy for the determination of butyrylcholinesterase (BChE) activity and organophosphorus (OPs) concentration. BChE promoted the hydrolysis of iodized s-butyrylthiocholine (BTCh) into a reducing substance thiocholine, which can decompose CoOOH nanosheets (CoOOH NSs) to Co2+. Subsequently, the single-stranded DNA (ssDNA) on the surface of CoOOH NSs was released. Then, ssDNA hybridized with hairpin DNA (h-DNA) and triggered the target recycling amplification process, producing large amounts of G-quadruplex. After adding thioflavin T (ThT), the target BChE was converted into activatable G-quadruplex/ThT with an amplified yellow fluorescence signal. The addition of OPs could significantly inhibit the hydrolysis of BTCh by BChE and thus unable to produce the yellow fluorescence G-quadruplex/ThT complex. Throughout the entire process, the fluorescence intensity of Hg-ZnSe QDs as a reference signal remained unchanged at 630 nm. Furthermore, this work provided an effective approach for detecting the BChE activity in serum samples and OPs in fruits and vegetables.

Accurate Capture and Identification of Exosomes: Nanoarchitecture of the MXene Heterostructure/Engineered Lipid Layer.

Recently, exosome detection has become an important breakthrough in clinical diagnosis. However, the effective capture and accurate identification of cancer exosomes in a complex biomatrix are still a tough task. Especially, the large size and non-conductivity of exosomes are not conducive to highly sensitive electrochemical or electrochemiluminescence (ECL) detection. Therefore, we have developed a Ti3C2Tx-Bi2S3-x heterostructure/engineered lipid layer-based nanoarchitecture to overcome the limitations. The engineered lipid layer not only specifically captured and efficiently fused CD63 positive exosomes but also showed excellent antifouling property in the biological matrix. Moreover, the MUC1 aptamer-modified Ti3C2Tx-Bi2S3-x heterostructure further identified and covered the gastric cancer exosomes that have been trapped in the engineered lipid layer. In the self-luminous Faraday cage-type sensing system, the Ti3C2Tx-Bi2S3-x heterostructure with sulfur vacancies extended the outer Helmholtz plane and amplified the ECL signal. Therefore, this sensor can be used to detect tumor exosomes in ascites of cancer patients without additional purification. It provides a new pathway to detect exosomes and other large-sized vesicles with high sensitivity.

A sensing platform for on-site detection of glutathione S-transferase using oxidized Pi@Ce-doped Zr-based metal-organic frameworks(MOFs).

The development of point-of-care testing (POCT) for glutathione S-transferase (GST) is an effective way to establish the mechanism of targeted monitoring of cancer chemotherapy drug metabolism. Assays for GST with high sensitivity as well as on-site screening have been urgently required to monitor this process. Herein, we synthesized oxidized Pi@Ce-doped Zr-based metal-organic frameworks (MOFs) by electrostatic self-assembly between phosphate and oxidized Ce-doped Zr-based MOFs. It was found that the oxidase-like activity of oxidized Pi@Ce-doped Zr-based MOFs was substantially increased after phosphate ion (Pi) assembly. And a stimulus-responsive hydrogel-based kit was constructed by embedding oxidized Pi@Ce-doped Zr-based MOFs into a PVA (polyvinyl alcohol) hydrogel system, we integrated a portable hydrogel kit with a smartphone for real-time monitoring of GST for quantitative and accurate analysis. The color reaction was triggered based on oxidized Pi@Ce-doped Zr-based MOFs with 3,3',5,5'-tetramethylbenzidine (TMB). However, in the presence of glutathione (GSH), the above color reaction was hindered due to the reducibility of GSH. Catalyzed by GST, GSH can react with 1-chloro-2,4-dinitrobenzo (CDNB) to form an adduct, which caused the color reaction to occur again, resulting in the color response of the kit. In combination with ImageJ software, the kit image information acquired by smartphone could be converted into hue intensity, providing a direct quantitative tool for the detection of GST with a detection limit of 0.19mU·L-1. Based on the advantages of simple operation and cost-effectiveness, the introduction of the POCT miniaturized biosensor platform will meet the requirements of on-site quantitative analysis of GST.

A dual-signal fluorometric and colorimetric sensing platform based on gold-platinum bimetallic nanoclusters for the determination of ß-galactosidase activity.

Herein, a novel dual-signal sensing system for the determination of ß-galactosidase (ß-Gal) activity was established, which was based on a dual-emission probe assembled from gold-platinum bimetallic nanoclusters (Au-Pt NCs) and rhodamine B. Under the catalysis of ß-Gal, 4-nitrophenyl ß-D-galactopyranoside (PNPG) was rapidly hydrolyzed to generate p-nitrophenol (PNP), which has an obvious UV absorption peak at 400 nm. The hydrolyzed product PNP can quench the fluorescence of Au-Pt NCs effectively by inner filter effect (IFE), and PNP had no impact on the fluorescence of rhodamine B, which will change the emission intensity ratio of Au-Pt NCs and rhodamine B. Therefore, the ratiometric fluorescent and colorimetric dual-signal sensor based on Au-Pt NCs and rhodamine B was successfully constructed for sensitive detection of ß-Gal activity. The linear detection range for the ratiometric fluorescence and colorimetric methods were 2.5-25 U/L and 15-55 U/L with detection limits of 1.2 U/L and 5.2 U/L, respectively. The developed assay method has been used for quantitative detection of ß-Gal in spiked serum samples and showed good performance. And the detection platform has high reliability and excellent selectivity, which opens a new avenue for the further application of Au-Pt NCs in chemical sensing and biological analysis.

Cascade reaction biosensor based on gold nanocluster decorated iron-cobalt oxide nanosheets as a superior peroxidase mimic for dual-mode detection of α-glucosidase and its inhibitor.

Herein, we have synthesized a novel kind of gold nanoclusters decorated iron-cobalt oxide nanosheets (His-AuNCs@FeCo-ONSs) assembled by electrostatic interaction, which possessed both outstanding peroxidase-like activity and fluorescence property. Taking advantage of our bifunctional hybrid nanozyme and enzyme cascade reactions, a sensitive dual-mode (colorimetric/fluorescent) detection method for α-glucosidase was constructed. The detection limits for α-glucosidase were 2.2 U/L and 3.3 U/L in fluorometric and colorimetric mode, respectively. This method not only provides high sensitivity, but also can correct itself to improve the accuracy of analysis due to the dual-response signals. Furthermore, it was employed for α-glucosidase determination in real samples and screening of α-glucosidase inhibitors.

A ratiometric fluorescence method based on nitrogen-doped carbon quantum dots for the determination of the activity of alkaline phosphatase.

Herein, a simple and sensitive ratiometric fluorescence sensing platform to detect alkaline phosphatase (ALP) activity is developed on the basis of yellow fluorescent nitrogen-doped carbon quantum dots (YNCDs). The hydrolysis of ascorbic acid 2-phosphate (AAP) into ascorbic acid (AA) is catalyzed by ALP. Then, AA will react with o-phenylenediamine (OPD) to form 3-(1,2-dihydroxyethyl)furo[3,4b]-quinoxaline (QXD) which is a blue fluorescent quinoxaline derivative with emission at 435 nm in the presence of Cu2+. YNCDs have yellow fluorescence emission at 555 nm, and can maintain stable in QXD reaction system. Therefore, by utilizing the fluorescence of YNCDs at 555 nm as reference signal and the fluorescence of QXD at 435 nm as report signal, we can detect the ALP activity by monitoring the fluorescence ratio (F435/F555). The linear range is 0.5-5 U/L, and the limit of detection is 0.14 U/L. An application of this method for the analysis of ALP in human serum has given satisfactory results. A ratiometric fluorescent nanoprobe for ascorbic acid and alkaline phosphatase detection with excellent biocompatible and high sensitivity was successfully constructed based on YNCDs and QXD.

Fluorescence sensing platform for sarcosine analysis based on nitrogen-doping copper nanosheets and gold nanoclusters.

Herein, nitrogen-doped copper nanosheet (CuT@N NS) was obtained by pyrolysis of the tryptophan-based Cu-containing nanosheet (CuT NS) and graphitic carbon nitride (g-C3N4). The high Cu loading (8.62 wt%) on the nitrogen-doped carbon-nanosheets endowed CuT@N NS with prominent peroxidase-mimicking activity. A fluorometric sensing platform comprising CuT@N NS with exceptional catalytic performance and 5-methyl-2-thiouracil capped gold nanoclusters (AuNCs) was successfully constructed for the determination of sarcosine. Under appropriate conditions, sarcosine oxidase (SOx) recognized and oxidized the sarcosine substrate to generate H2O2. The as-obtained CuT@N NS then decomposed H2O2 into superoxide radicals (O2•-) and further promoted the reaction between 4-aminoantipyrine (AAP) and phenol to form pink-red quinone-imine dye (p-QID, absorbance at 508 nm). When excited at 380 nm, the orange-emitting AuNCs peak at 560 nm was quenched by the produced p-QID via inner filter effect (IFE). Thus, a CuT@N NS/AuNCs-based fluorometric sensing strategy for sarcosine analysis was constructed. Impressively, the outputting fluorometric signal revealed a wide linear range of 7.5-1100 µmol L-1 with the detection limit of 4.69 µmol L-1. These findings not only expanded the applications of nanozyme in bioanalysis, but also provided a sensitive and effective approach for monitoring the levels of sarcosine.