Tetramethylbenzidine Research Articles

Luminescent gold nanoclusters loaded on iron metal–organic framework with enhanced peroxidase-like activity for the colorimetric/fluorescent determination of H2O2

In this work, a composite nanozyme, termed AuNCs/Fe-MIL, was obtained by combining glutathione-stabilized gold nanoclusters (AuNCs) on the surface of the Fe metal–organic framework (Fe-MIL-88B-NH2). The AuNCs/Fe-MIL nanocomposite retains the red emission of AuNCs, exhibiting unique dual-emission fluorescence with remarkable stability. This nanozyme exhibits improved peroxidase-like activity as a result of the heightened rate of electron transfer. As a result, the decomposition of H2O2 is accelerated, leading to the generation of more •OH radicals that oxidize 3,3′,5,5′-tetramethylbenzidine (TMB) into blue ox-TMB. The generated ox-TMB can quench the fluorescence of AuNCs/Fe-MIL in the red-light region. By leveraging the dual function of the catalytic product ox-TMB, a fusion of colorimetric and fluorescence signals enabled H2O2 detection within 10 min. The sensing platform demonstrated wide linear ranges of 5–1000 μM with a detection limit of 2.78 μM in colorimetry mode and 5–500 μM with a detection limit of 0.78 μM in fluorescence mode. This method was successfully utilized for accurate in situ monitoring of H2O2 release from different types of living cells. Additionally, a dual-mode portable sensing platform based on AuNCs/Fe-MIL/TMB hydrogel was constructed for convenient H2O2 detection. Through a smartphone application, precise quantitation of H2O2 added to milk samples was achieved. This work provides a rational design of multifunctional nanozymes for reliable biomimetic catalysis.

The Oxidase Mimicking Activity of MnOx NPs/Co3O4 NPs Hybrid Nanozyme for Glucose Oxidation

Herein, the hybrid nanozyme MnOx NPs/Co3O4 NPs on indium tin oxide coated glass substrate (ITO) was manufactured by imparting the porous morphology with its distinct merits: its surface valence states, oxygen vacancies, large surface area, and abundant active sites. The oxidase-like activity was investigated via the catalytic oxidation of chromogenic substrate in the presence of glucose visualized by the eyes. MnOx NPs containing Mn2+ and Mn3+ have a superior ability to oxidize glucose by reducing dissolved oxygen and producing H2O2. Co3O4 NPs, in turn, reduce H2O2 with concomitant 3,3′,5,5′-tetramethylbenzidine (TMB) oxidization. Thus, the nanozyme mimics the dual roles of glucose oxidase and peroxidase. The oxidase-like activity of hybrid nanozyme for glucose was found to be higher than those of single components. The nanozyme responded to glucose with a linear range from 60 µM to 1200 μM. The acceptable performance is probably due to the facilitated access of glucose to the proximity of the sensor surface. Good reproducibility was accomplished by virtue of the meticulous construction of NPs. Without functionalization and enzyme utilization, the fabricated nanozyme holds promise as a substitute for peroxidase and oxidase for detecting glucose.

Reliable ratiometric colorimetric monitoring of dopamine in practice based on the catalytic signal amplification of nano CeO2/CuO modified carboxylated chitosan

To reliably monitor dopamine (DA), a common neurotransmitter, a kind of nano CeO2/CuO modified carboxylated chitosan nanozyme (CeO2/CuO@CC) was constructed under microwave-assisted conditions. The proposed CeO2/CuO@CC exhibited superior synergistic- enhancement oxidase-like activity, with high specific surface area and superior stability. It could accelerate the oxidization of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into blue oxTMB in the air. What’s more, the presence of trace DA could exclusively change the enzyme-like activity of CeO2/CuO@CC, accompanied with visual color changes from blue to colorless. Under the optimized conditions, there expressed double ratiometric linear relationships between lg(A652/A285)/lg(A370/A285) and cDA, with a large linear range between 6.0 × 10−9 and 1.0 × 10−5 M of DA. When applied for visual detection of DA in real samples, the detection limit was low to 2.0 × 10−9 M with RSD % less than 4.14 %. The mechanisms for enhanced oxidase-mimic activity of CeO2/CuO@CC and its selective detection of DA were proposed. This work will offer an insight to design efficient nanozyme for visual detection of bio-analytes in practice.

Phosphorothioate-modified G-quadruplex as a signal-on dual-mode reporter for CRISPR/Cas12a-based portable detection of environmental pollutants

BackgroundClustered regularly interspaced short palindromic repeats (CRISPR)/Cas12a-powered biosensor with a G-quadruplex (G4) reporter offer the benefits of simplicity and sensitivity, making them extensively utilized in detection applications. However, these biosensors used for monitoring pollutants in environmental water samples may face the problem of high background signal and easy interference due to the “signal-off” output. It is obvious that a biosensor based on the CRISPR/Cas12a system and G4 with a “signal on” output mode needs to be designed for detecting environmental pollutants. ResultsBy using phosphorothioate-modified G4 as a reporter and catalytic hairpin assembly (CHA) integrated with Cas12a as an amplification strategy, a “signal-on” colorimetric/photothermal biosensor (psG4-CHA/Cas) for portable detection of environmental pollutants was developed. With the help of functional nucleotides, the target pollutant (kanamycin or Pb2+) triggers a CHA reaction to produce numerous double-strand DNA, which can activate Cas12a′s trans-cleavage activity. The active Cas12a cleaves locked DNA to release caged psG-rich sequences. Upon binding hemin, the psG-rich sequence forms a psG4/hemin complex, facilitating the oxidation of the colorless 3,3′,5,5′-tetramethylbenzidine (TMB) into the blue photothermal agent (oxTMB). The smartphone was employed for portable colorimetric detection of kanamycin and Pb2+. The detection limits were found to be 100 pM for kanamycin and 50 pM for Pb2+. Detection of kanamycin and Pb2+ was also carried out using a portable thermometer with a detection limit of 10 pM for kanamycin and 8 pM for Pb2+. SignificanceSensitive, selective, simple and robust detection of kanamycin and Pb2+ in environmental water samples is achieved with the psG4-CHA/Cas system. This system not only provides a new perspective on the development of efficient CRISPR/Cas12a-based “signal-on” designs, but also has a promising application for safeguarding human health and environmental monitoring.

A dual-mode fluorometric/colorimetric sensor for sulfadimethoxine detection based on Prussian blue nanoparticles and carbon dots.

A dual-mode (colorimetric/fluorescence) nanoenzyme-linked immunosorbent assay (NLISA) was developed based on Au-Cu nanocubes generating Prussian blue nanoparticles (PBNPs). It is expected that this method can be used to detect the residues of sulfonamides in the field, and solve the problem of long analysis time and high cost of the traditional method. Sulfadimethoxine (SDM) was selected as the proof-of-concept target analyte. The Au-Cu nanocubes were linked to the aptamer by amide interaction, and the Au-Cu nanocubes, SDM and antibody were immobilized on a 96-well plate using the sandwich method. The assay generates PBNPs by oxidising the Cu shells on the Au-Cu nanocubes in the presence of hydrochloric acid, Fe3+ and K3[Fe (CN)6]. In this process, the copper shell undergoes oxidation to Cu2+ and subsequently Cu2 + further quenches the fluorescence of the carbon point. PBNPs exhibit peroxidase-like activity, oxidising 3,3',5,5'-tetramethylbenzidine (TMB) to OX-TMB in the presence of H2O2, which alters the colorimetric signal. The dual-mode signals are directly proportional to the sulfadimethoxine concentration within the range 10- 3~10- 7 mg/mL. The limit of detection (LOD) of the assay is 0.023 ng/mL and 0.071 ng/mL for the fluorescent signal and the colorimetric signal, respectively. Moreover, the assay was successfully applied to determine sulfadimethoxine in silver carp, shrimp, and lamb samples with satisfactory results.

Silicene's intriguing nanozymatic activity: Improved colorimetric and electrochemically supported colorimetric biosensing

Silicene is a material that is largely studied via theoretical models. In the practical world, lengthy incubation times and poor sensitivity of colorimetric detection methods have limited their implementation. This paper transforms silicene's theoretical advantages into practical applications, presenting it as a tangible solution for significantly improving colorimetric biosensing. More specifically, this paper presents a pivotal breakthrough by not only documenting the remarkable nanozymatic activity of silicene in oxidizing 3,3′,5,5′-Tetramethylbenzidine (TMB) with hydrogen peroxide (H2O2), but also leveraging silicene’s inherent electronic properties to enhance the speed and precision of the peroxidase reaction, thereby improving its performance and advancing applicability. Two detection mechanisms were tested to observe the influence of silicene’s electronic scope; one in the absence and the other in the presence of an externally supplied electric potential. Both were successful in the detection of H2O2 and dopamine. The latter proved to be more favorable as it not only shortened the incubation time from 30 min to 1.5 min but also exhibited lower Limit of Detection (LOD) values. To demonstrate, for dopamine, in a concentration range of 0 – 10 μM, the LOD was 4.13 μM without the electric potential and 2.21 μM with the electric potential. Similarly, in a second concentration range, the LOD values were 37.81 μM and 34.94 μM, respectively. Kinetic profiles indicated the superior affinity silicene has for TMB and H2O2. A selectivity test also indicated that the method has excellent selectivity towards dopamine.

Platinum-nickel nanoparticle-based oxidase-like nanozyme for colorimetric/photothermal dual-mode detection of organophosphorus pesticides

In this study, based on the oxidase-like (OXD) activity of platinum-nickel nanoparticles (Pt-Ni NPs) nanozyme and further in collaboration with 3,3′,5,5′-tetramethylbenzidine (TMB), a colorimetric/photothermal dual-mode probe for organophosphorus pesticides (OPs) detection was developed. TMB undergone the catalytic oxidation of Pt-Ni NPs to change from colorless to blue oxidized substance (oxTMB) and exhibited higher temperature under near-infrared irradiation. The hydrolysis product of acid phosphatase (ACP) with L-ascorbic acid 2-phosphate trisodium competed with the OXD-like activity of Pt-Ni NPs to prevent the oxidation of TMB. As the concentration of OPs grew, inhibiting the blocking effect by deactivating ACP, the generated oxTMB caused notable color and temperature changes in the reaction system. The dual-mode probe responded linearly to chlorpyrifos, with the detection limits for colorimetric and photothermal modes of 1.2 ng/mL and 1.66 ng/mL, respectively. This work proposes a simple and reliable colorimetric/photothermal platform for OPs identification, which overcomes the problem that single-mode detection probes are susceptible to external factors, and has broad application potential in the field of food safety.

All-in-one smartphone imaging for visual immunobiosensing of cancer biomarker using electronic gyro-centrifuge and nanozyme

Integrated functionality that combines recognition elements and sensors on a single platform to provide real-time detection capabilities for fifth-generation biosensors is considered one of the dominant tools for addressing point-of-care testing in the community today. Inspired by gyroscope and smartphone-based sensors, we developed a portable and low-cost immunobiosensor for high-throughput detection of cancer-related markers. The main testing process was divided into two parts: blood sample preprocessing based on an electronic gyroscope and signal reading based on nanozyme and smartphone. The feasibility of a commercial electronic gyroscope for serum acquisition from whole blood samples was evaluated. In addition, PtCd nanoclusters (PtCd NCs) were used as excellent peroxidase-like nanozymes to trigger the target signal conversion of the immunosensor. PtCd NCs were able to catalyze the conversion of H2O2 to ·OH, which rapidly oxidized the colorless 3,3′,5,5′-tetramethylbenzidine (TMB) substrate to the blue Oxidized-TMB (oxTMB). Naked eye visible obvious color change and the RGB values of the color metrology system could also be read by smartphone. Prostate-specific antigen (PSA) as a model biomarker was used to validate the utility of an integrated immunoassay system to enable early cancer screening through localization of RGB values. This work focuses on convenient blood sample preprocessing and simplified serologic cancer screening steps to develop an efficient and cost-effective immunoassay platform for community and on-site point-of-care testing (POCT) in resource-limited settings.

Graphene quantum dots on TiO2 nanotubes as a light-assisted peroxidase nanozyme

Hybrid nanozyme graphene quantum dots (GQDs) deposited TiO2 nanotubes (NTs) on titanium foil (Ti/TiO2 NTs-GQDs) were manufactured by bestowing the hybrid with the advantageous porous morphology, surface valence states, high surface area, and copious active sites. The peroxidase-like activity was investigated through the catalytic oxidation of chromogenic substrate 3,3′,5,5′-tetramethylbenzidine (TMB) in the presence of H2O2, which can be visualized by the eyes. TiO2 NTs and GQDs comprising oxygen-containing functional groups can oxidize TMB in the presence of H2O2 by mimicking peroxidase enzymes. The peroxidase-mimicking activity of hybrid nanozyme was significantly escalated by introducing light illumination due to the photosensitive features of the hybrid material. The peroxidase-like activity of Ti/TiO2 NTs-GQDs enabled H2O2 determination over the linear range of 7 to 250 μM, with a LOD of 2.1 µM. The satisfying peroxidase activity is possibly due to the unimpeded access of H2O2 to the catalyst’s active sites. The porous morphology provides the easy channeling of reactants and products. The periodic structure of the material also gave rise to acceptable reproducibility. Without material functionalization, the Ti/TiO2 NTs-GQDs can be a promising substitute for peroxidases for H2O2 detection.Graphical abstract

MOF-based Ag NPs/Co3O4 nanozyme for colorimetric detection of thiophanate-methyl based on analyte-enhanced sensing mechanism.

Silver nanoparticles supported on metal-organic framework (ZIF-67)-derived Co3O4 nanostructures (Ag NPs/Co3O4) were synthesized via a facile in situ reduction strategy. The resulting materials exhibited pH-switchable peroxidase/catalase-like catalytic activity. Ag NP doping greatly enhanced the catalytic activity of Ag NPs/Co3O4 towards 3,3',5,5'-tetramethylbenzidine (TMB) oxidation and H2O2 decomposition which were 59 times (A652 of oxTMB) and 3 times (A240 of H2O2) higher than that of ZIF-67, respectively. Excitingly, thiophanate-methyl (TM) further enhanced the peroxidase-like activity of Ag NPs/Co3O4 nanozyme due to the formation of Ag(I) species in TM-Ag NPs/Co3O4 and generation of more radicals resulting from strong interaction between Ag NPs and TM. The TM-Ag NPs/Co3O4 nanozyme exhibited lower Km and higher Vmax values towards H2O2 when compared with Ag NPs/Co3O4 nanozyme. A simple, bioelement-free colorimetric TM detection method based on Ag NPs/Co3O4 nanozyme via analyte-enhanced sensing strategy was successfully established with high sensitivity and selectivity. Our study demonstrated that hybrid noble metal NPs/MOF-based nanozyme can be a class of promising artificial nanozyme in environmental and food safety applications.

Synergistic signal amplification of ultrathin nanowire-like PtCoFeRuMo high-entropy nanozyme and Z-scheme WO3/ZnIn2S4 heterostructure for split-typed photoelectrochemical aptasensing of myoglobin

Enzyme-initiated sensing strategy has wide applications in photoelectrochemical (PEC) analysis for its high specificity and excellent catalytic efficiency. Herein, ultrathin PtCoFeRuMo high-entropy alloy nanowires (PtCoFeRuMo HEANWs) were prepared by a simple solvothermal method, whose peroxidase-like (POD-like) catalytic property was critically investigated by tetramethylbenzidine (TMB) oxidation in the presence of H2O2. Simultaneously, Z-scheme WO3/ZnIn2S4 heterostructures (labeled WO3/ZnIn2S4) were constructed under solvothermal treatment, whose optical properties and photoactivity were rigorously studied by a set of techniques, combined by elaborating the interfacial charge transfer mechanism. On such foundation, a split-typed PEC aptasensor was established with the WO3/ZnIn2S4 substrate, whose detection signals were further magnified through catalytic oxidation of 3-amino-9-ethylcarbazole (AEC) initiated by the PtCoFeRuMo HEANWs. The resultant PEC sensor realized excellent bioanalysis of biomarker myoglobin, showing a broader linear range from 1.0 × 10−2 to 1.0 × 105 pg mL−1 and a lower limit of detection (0.96 fg mL−1, S/N = 3). This split-typed PEC biosensing strategy shows substantial promise in bioanalysis and clinical diagnosis.

Lab on a bead with oscillatory centrifugal microfluidics for fast and complete mixing enables fast and accurate biomedical assays

Rapid mixing and precise timing are key for accurate biomedical assay measurement, particularly when the result is determined as the rate of a reaction: for example rapid immunoassay in which the amount of captured target is kinetically determined; determination of the concentration of an enzyme or enzyme substrate; or as the final stage in any procedure that involves a capture reagent when an enzyme reaction is used as the indicator. Rapid mixing and precise timing are however difficult to achieve in point-of-care devices designed for small sample volumes and fast time to result. By using centrifugal microfluidics and transposing the reaction surface from a chamber to a single mm-scale bead we demonstrate an elegant and easily manufacturable solution. Reagents (which may be, for example, an enzyme, enzyme substrate, antibody or antigen) are immobilised on the surface of a single small bead (typically 1–2 mm in diameter) contained in a cylindrical reaction chamber subjected to periodically changing rotational accelerations which promote both mixing and uniform mass-transfer to the bead surface. The gradient of Euler force across the chamber resulting from rotational acceleration of the disc, dΩdisc/dt, drives circulation of fluid in the chamber. Oscillation of Euler force by oscillation of rotational acceleration with period, T, less than that of the hydrodynamic relaxation time of the fluid, folds the fluid streamlines. Movement of the bead in response to the fluid and the changing rotational acceleration provides a dynamically changing chamber shape, further folding and expanding the fluid. Bead rotation and translation driven by fluid flow and disc motion give uniformity of reaction over the surface. Critical parameters for mixing and reaction uniformity are the ratio of chamber radius to bead radius, rchamber/rbead, and the product Trchamber(dΩdisc/dt), of oscillation period and Euler force gradient across the fluid. We illustrate application of the concept using the reaction of horse radish peroxidase (HRP) immobilised on the bead surface with its substrate tetramethylbenzidine (TMB) in solution. Acceleration from rest to break a hydrophobic valve provided precise timing for TMB contact with the bead. Solution uniformity from reaction on the surface of the bead in volumes 20–50 uL was obtained in times of 2.5 s or less. Accurate measurement of the amount of surface-bound HRP by model fitting to the measured kinetics of colour development at 10 s intervals is demonstrated.

Realizing Ultrasensitive and Accurate Point-of-Care Profiling for ATP with a Triple-Mode Strategy Based on the ATP-Induced Reassembly of a Copper Coordination Polymer Nanoflower.

New strategies for accurate and reliable detection of adenosine triphosphate (ATP) with portable devices are significant for biochemical analysis, while most recently reported approaches cannot satisfy the detection accuracy and independent of large instruments simultaneously, which are unsuitable for fast, simple, and on-site ATP monitoring. Herein, a unique, convenient, and label-free point-of-care sensing strategy based on novel copper coordination polymer nanoflowers (CuCPNFs) was fabricated for multimode (UV-vis, photothermal, and RGB values) onsite ATP determination with high selectivity, sensitivity, and accuracy. The resulting CuCPNFs with a 3D hierarchical structure exhibit the ATP-triggered decomposition behavior because the competitive coordination between ATP and the copper ions of CuCPNFs can result in the formation of ATP-Cu, which reveals preeminent peroxidase mimics activity and can accelerate the oxidation of 3, 3', 5, 5'-tetramethylbenzidine (TMB) to form oxTMB. During this process, the detection system displayed not only color changes but also a strong NIR laser-driven photothermal effect. Thus, the photothermal and color signal variations are easily monitored by a portable thermometer and a smartphone. This multimode point-of-care platform can meet the requirements of onsite, without bulky equipment, accuracy, and reliability all at once, greatly enhancing its application in practice and paving a new way in ATP analysis.

A fluorescence and colorimetric dual-mode immunoassay for detection of ochratoxin A based on cerium nanoparticles

Ochratoxin A (OTA) is a widely distributed mycotoxin that seriously endangers food safety and human health. Therefore, establishing a sensitive and simple method for detecting OTA is crucial to ensure food safety. A fluorescence and colorimetric dual-mode immunoassay was established using cerium-based nanoparticles CPNs(Ⅳ) for detection of OTA. CPNs(IV) with good oxidase activity can catalyze the oxidation of colorless 3,3′,5,5′-tetramethylbenzidine (TMB) to produce visible blue color signal. Under the catalysis of ALP, the ascorbic acid 2-phosphate (AA2P) was hydrolyzed to form ascorbic acid (AA), which reduced CPNs(Ⅳ) to CPNs(III). The produced CPNs(III) can emit significant fluorescence signal. Because the concentration of OTA was inversely proportional to the concentration of IgG-ALP in the microplate, the fluorescence signal was gradually decreased and the blue color signal gradually increased with the addition of OTA. The limits of detection (LODs) of this dual-mode immunoassay were 0.404 ng/mL (for fluorescent mode) and 0.962 ng/mL (for colorimetric mode). The recoveries of OTA were 97.60 %–103.55 % for the fluorescence mode and 99.12 %–102.60 % for the colorimetric mode in corn samples. These results indicated the dual-mode immunoassay was sensitive and reliable for detection of OTA in actual samples.

Colorimetric and Electrochemical Dual-Mode Detection of Thioredoxin 1 Based on the Efficient Peroxidase-Mimicking and Electrocatalytic Property of Prussian Blue Nanoparticles

As a potent detection method for cancer biomarkers in physiological fluid, a colorimetric and electrochemical dual-mode sensing platform for breast cancer biomarker thioredoxin 1 (TRX1) was developed based on the excellent peroxidase-mimicking and electrocatalytic property of Prussian blue nanoparticles (PBNPs). PBNPs were hydrothermally synthesized using K3[Fe(CN)6] as a precursor and polyvinylpyrrolidone (PVP) as a capping agent. The synthesized spherical PBNPs showed a significant peroxidase-like activity, having approximately 20 and 60% lower Km values for 3,3′,5,5′-tetramethylbenzidine (TMB) and H2O2, respectively, compared to those of horseradish peroxidase (HRP). The PBNPs also enhanced the electron transfer on the electrode surface. Based on the beneficial features, PBNPs were used to detect target TRX1 via sandwich-type immunoassay procedures. Using the strategies, TRX1 was selectively and sensitively detected, yielding limit of detection (LOD) values as low as 9.0 and 6.5 ng mL−1 via colorimetric and electrochemical approaches, respectively, with a linear range of 10–50 ng mL−1 in both strategies. The PBNP-based TRX1 immunoassays also exhibited a high degree of precision when applied to real human serum samples, demonstrating significant potentials to replace conventional HRP-based immunoassay systems into rapid, robust, reliable, and convenient dual-mode assay systems which can be widely utilized for the identification of important target molecules including cancer biomarkers.

Au/Ag2MoO4 Nanocomposite: A Dual-Function Catalyst for Dye Degradation and Colorimetric Detection

The authors have successfully synthesized a distinctive potato-shaped Au/Ag2MoO4 nanocomposite with exceptional photocatalytic capabilities in the degradation of methylene blue (MB) under UV irradiation even in neutral conditions. Furthermore, this material displays noteworthy peroxidase-like oxidation activity, facilitating the detection of hydrogen peroxide (H2O2) and ascorbic acid (AA) in aqueous environments. This detection method relies on the oxidation of 3,3',5,5'-tetramethylbenzidine (TMB). The signal generated by TMB adsorption on the catalyst's surface plays a pivotal role in the oxidation of TMB in the presence of H2O2, resulting in the formation of a distinct blue-colored product. This reaction is equally influenced by the presence of AA, which, owing to its antioxidant properties, impedes the development of the color signal. This colorimetric approach exhibits high sensitivity, boasting low limits of detection (LOD) and quantification (LOQ) of 0.0252 and 0.0839mM for H2O2, and 1.7724 and 5.7109mg/L for AA, respectively. The method offers a broad linear range spanning from 0.029 to 26.47mM for H2O2 and 0 to 40mg/L for AA with a straightforward, cost-effective analytical procedure, and yields highly reliable results. The proposed method displays superior selectivity in detecting AA and high anti-interference capabilities. These findings underscore the photocatalytic and peroxidation-like activities of Au/Ag2MoO4 nanocomposite, positioning it as a promising candidate for various biomedical applications.

Near-infrared Photoelectrochemical and colorimetric dual-mode aptasensor for determining osteopontin based on Er-MOF nanoballs@gold nanoparticles and Au2Pt nanozymes

A near infrared photoelectrochemical (PEC) and colorimetric dual-mode aptasensor was fabricated for efficiently determining osteopontin (OPN). Er-MOFs and Au2Pt nanoparticles were successfully synthesized. The gold nanoparticles (AuNPs) modified Er-MOF (Er-MOFs@AuNPs) nanocomposites were introduced into the glassy carbon electrode (GCE) to construct a photoelectrochemical sensing interface. OPN aptamer (Apt1) was immobilized onto the Er-MOFs@AuNPs based photoelectrochemical platform through Au-S bond for the selectively recognizing osteopontin. Au2Pt-labeled OPN aptamer (Au2Pt-Apt2) was subsequently captured onto the OPN/Apt1/MOFs@AuNPs/GCE surface to form a sandwich sensing interface, leading to an enhancement on the photocurrent response. Interestingly, Au2Pt nanozyme which possesses peroxidase activity could produce a solution color change in the presence of 3,3',5,5'-tetramethylbenzidine (TMB) and hydrogen peroxide. The variations of photocurrent response and absorbance can be employed for the quantitative detection of OPN. Under the optimal conditions, the PEC and colorimetric sensing strategies show linear responses towards OPN in the concentration range of 0.01 ~ 25ngmL-1 and 1 ~ 850ngmL-1, respectively. The calculated determination limits were 2.6pgmL-1 and 0.04ngmL-1. The PEC and colorimetric dual-mode sensing platform can provide no crosstalk results and can be mutually verified, thus effectively improving the accuracy and reliability for the determination of OPN.

CeO2 nanocages with tetra-enzyme mimetic activities for dual-channel ratiometric colorimetric detection of microcystins-LR

BackgroundMicrocystin-leucine-arginine (MC-LR) produced by various cyanobacteria during harmful algal bloom poses serious threats to drinking water safety and human health. Conventional chromatography-based detection methods require expensive instruments and complicated sample pretreatment, limiting their application for on-site detection. Colorimetric aptasensors are simple and rapid, and are amenable to fast detection. However, they provide only one output signal, resulting in poor sensitivity and accuracy. Dual-channel ratiometric colorimetric method based on the peroxidase-like activity of nanozyme can achieve self-calibration by recording two reverse signals, providing significantly enhanced sensitivity and accuracy. ResultsCeO2 nanocages (CeO2 NCs) with tetra-enzyme mimetic activities (oxidase-, peroxidase-, catalase- and superoxide dismutase-like activities) were facilely synthesized using zeolitic imidazolate framework-67 (ZIF-67) as sacrificial template. The peroxidase-like activity of CeO2 NCs can be regulated by DNA, and it showed opposite response to two chromogenic substrates (2,2’-azino-bis-(3-ethylbenzthiazoline-6-sulfonic acid) (ABTS) and 3,3',5,5'-tetramethylbenzidine (TMB)), which was mainly attributed to the changed affinity. On the basis of MC-LR aptamer-tunable peroxidase-like activity of CeO2 NCs in TMB and ABTS channel, a dual-channel ratiometric colorimetric aptasensor was constructed for detection of MC-LR. Compared with conventional single-signal colorimetric assays, the proposed method showed lower limit of detection (0.66 pg mL−1) and significantly enhanced sensitivity. Moreover, the practicability of the ratiometric colorimetric assay was demonstrated by detecting MC-LR in real water samples, and satisfactory recoveries (94.9-101.9%) and low relative standard deviations (1.6-6.3%) were obtained. SignificanceThis work presents a nanozyme-based ratiometric colorimetric aptasensor for MC-LR detection by recording the reverse responses of two chromogenic reactions. Benefiting from the self-calibration function, the method can achieve higher sensitivity and accuracy. The short detection time and practical application in real water samples show greating potential for environmental monitoring.

A Smartphone-Enabled Colorimetric Platform Based on Enzyme Cascade Amplification Strategy for Detection of Staphylococcus aureus in Milk

Staphylococcus aureus (S. aureus) is a pathogenic bacterium-contaminating milk and dairy foods causing food poisoning and foodborne pathogens. In this work, a smartphone-enabled enzyme cascade-triggered colorimetric platform was constructed using cascade bio-nanozyme formed by immobilized glucose oxidase (GOx) on the Fe3O4@Ag for rapid detection of S. aureus. Benefiting from reasonable experimental design, a bio-nanozyme cascade-triggered reaction was achieved through H2O2 produced by GOx oxidation of glucose, followed by in situ catalysis of 3,3',5,5'-tetramethylbenzidine (TMB) by the inherent peroxidase-like activity of Fe3O4@Ag to produce color signals. S. aureus detection could be performed through naked-eye observation and smartphone measurement, the developed assay can achieve quantitative and qualitative detection of S. aureus. The on-site nanoplatform had satisfactory specificity and sensitivity with a low detection limit of 6.9 cfu·mL-1 in 50 min. Moreover, the nanoplatform has good practicality in the detection of S. aureus in milk samples. Therefore, the assay has potential application prospects in food safety inspection.

Colorimetric aptasensor based on magnetic beads and gold nanoparticles for detecting mucin 1

In this work, a colorimetric aptasensor based on magnetic beads (MBs), gold nanoparticles (AuNPs) and Horseradish Peroxidase (HRP) was prepared for the detection of mucin 1 (MUC1). Complementary DNA of the MUC1 aptamer (Apt) immobilized on the MBs was combined with the prepared AuNPs-Apt-HRP complex (AuNPs@Apt-HRP). In the presence of MUC1, it specifically bound to Apt, resulting in the detachment of gold nanoparticles from the MBs. After magnetic separation, AuNPs@Apt-HRP was separated into the supernatant and reacted with 3,3′,5,5′-Tetramethylbenzidine (TMB) to produce color reaction from colorless to blue. The linear range of MUC1 was from 75 to 500 μg/mL (R2 = 0.9878), and the detection limit was 41.95 μg/mL. The recovery rate of MUC1 in human serum was 99.18 %∼101.15 %. This method is simple and convenient. Moreover, it does not require complex and expensive equipment for detection of MUC1. It provides value for the development of MUC1 colorimetric sensors and a promising strategy for the determination of MUC1 in clinical diagnosis.