Phosphatase-mimicking Zr@PDA nanozyme with excellent dispersion stability for the detection of fructose 1,6-diphosphate.

Zr4+-doped polydopamine (Zr@PDA) nanozyme with phosphatase-like activity was synthesized by a one-pot hydrothermal method for the first time. Compared with previous representative phosphatase-mimicking nanozymes (i.e., CeO2 NPs, ZrO2 NPs and UiO-66), Zr@PDA not only exhibited higher dispersion stability in water, but also higher catalytical efficiency. Kcat/Km of Zr@PDA is 35 and 12 times that of UiO-66 and ZrO2 NPs, respectively, which would endow the Zr@PDA-based analytical methods with high sensitivity. As a demonstration, a novel colorimetric method based on Zr@PDA nanozyme was developed for sensitive detection of the drug fructose 1,6-diphosphate. The linear range is 1-15 µM with a detection limit as low as 0.38 µM.

Scaling up of a Self-Confined Catalytic Hybridization Circuit for Robust microRNA Imaging.

The precise regulation of cellular behaviors within a confined, crowded intracellular environment is highly amenable in diagnostics and therapeutics. While synthetic circuitry system through a concatenated chemical reaction network has rarely been reported to mimic dynamic self-assembly system. Herein, a catalytic self-defined circuit (CSC) for the hierarchically concatenated assembly of DNA domino nanostructures is engineered. By incorporating pre-sealed symmetrical fragments into the preying hairpin reactants, the CSC system allows the hierarchical DNA self-assembly via a microRNA (miRNA)-powered self-sorting catalytic hybridization reaction. With minimal strand complexity, this self-sustainable CSC system streamlined the circuit component and achieved localization-intensified cascaded signal amplification. Profiting from the self-adaptively concatenated hybridization reaction, a reliable and robust method has been achieved for discriminating carcinoma tissues from the corresponding para-carcinoma tissues. The CSC-sustained self-assembly strategy provides a comprehensive and smart toolbox for organizing various hierarchical DNA nanostructures, which may facilitate more insights for clinical diagnosis and therapeutic assessment.

Boron nanosheets as a phosphatase mimicking nanozyme with ultrahigh catalytic activity for prodrug-based cancer therapy.

Boron nanosheets (BNSs) are reported as a new phosphatase mimicking nanozyme. Surprisingly, the catalytic rate of BNSs is up to 17 times those of known phosphatase mimicking nanozymes. By adding polyols and Lewis bases, the catalytic activity of BNSs was attributed to the Lewis acidity of the B centers of the BNSs. Theoretical investigation shows that the B centers are responsible for the catalytic hydrolysis of phosphoesters. Moreover, the biomimetic activity of the BNSs was further explored for enhancing anticancer therapy through nanozyme-catalyzed prodrug conversion.

Accelerating the Phosphatase-like Activity of Uio-66-NH2 by Catalytically Inactive Metal Ions and Its Application for Improved Fluorescence Detection of Cardiac Troponin I.

Compared with natural enzymes, nanozymes usually exhibit much lower catalytic activities, which limit the sensitivities of nanozyme-based immunoassays. Herein, several metal ions without enzyme-like activities were engineered onto Uio-66-NH2 nanozyme through postsynthetic modification. The obtained Mn+@Uio-66-NH2 (Mn+ = Zn2+, Cd2+, Co2+, Ca2+and Ni2+) exhibited improved phosphatase-like catalytic activities. In particular, a 12-fold increase in the catalytic efficiency (kcat/Km) of Uio-66-NH2 was observed after the modification with Zn2+. Mechanism investigations indicate that both the amino groups and oxygen-containing functional groups in Uio-66-NH2 are the binding sites of Zn2+, and the modified Zn2+ ions on Uio-66-NH2 serve as the additional catalytic sites for improving the catalytic performance. Furthermore, the highly active Zn2+@Uio-66-NH2 was used as a nanozyme label to develop a fluorescence immunoassay method for the detection of cardiac troponin I (cTnI). Compared with pristine Uio-66-NH2, Zn2+@Uio-66-NH2 can widen the linear range by 1 order of magnitude (from 10 pg/mL-1 µg/mL to 1 pg/mL-1 µg/mL) and also lower the detection limit by 5 times (from 4.7 pg/mL to 0.9 pg/mL).

Engineering of a Self-Regulatory Bidirectional DNA Assembly Circuit for Amplified MicroRNA Imaging.

The engineering of catalytic hybridization DNA circuits represents versatile ways to orchestrate a complex flux of molecular information at the nanoscale, with potential applications in DNA-encoded biosensing, drug discovery, and therapeutics. However, the diffusive escape of intermediates and unintentional binding interactions remain an unsolved challenge. Herein, we developed a compact, yet efficient, self-regulatory assembly circuit (SAC) for achieving robust microRNA (miRNA) imaging in live cells through DNA-templated guaranteed catalytic hybridization. By integrating the toehold strand with a preblocked palindromic fragment in the stem domain, the proposed miniature SAC system allows the reactant-to-template-controlled proximal hybridization, thus facilitating the bidirectional-sustained assembly and the localization-intensified signal amplification without undesired crosstalk. With condensed components and low reactant complexity, the SAC amplifier realized high-contrast intracellular miRNA imaging. We anticipate that this simple and template-controlled design can enrich the clinical diagnosis and prognosis toolbox.

Few-layered boron nitride nanosheet as a non-metallic phosphatase nanozyme and its application in human urine phosphorus detection.

Human urine phosphorus (existing in the form of phosphate) is a biomarker for the diagnosis of several diseases such as kidney disease, hyperthyroidism, and rickets. Therefore, the selective detection of phosphate in urine samples is crucial in the field of clinical diagnosis. Herein, we reported the phosphatase-like catalytic activity of few-layered h-BNNS for the first time. As the phosphatase-like activity of few-layered h-BNNS could be effectively inhibited by phosphate, a selective fluorescent method for the detection of phosphate was proposed. The linear range for phosphate detection is 0.5-10 µM with a detection limit of 0.33 µM. The fluorescent method was then explored for the detection of human urine phosphorus in real samples. The results obtained by the proposed method were consistent with those of the traditional method, indicating that the present method has potential application for urine phosphorus detection in clinical disease diagnosis.

Ratiometric fluorescent detection of total phosphates in frozen shrimp samples using catalytic active Zr(IV) modified gold nanoclusters.

Phosphate salts are important food additives in a variety of foods. In this study, the Zr(IV) modified gold nanoclusters (Au NCs) were prepared for ratiometric fluorescent sensing of phosphate additives in seafood samples. Compared with bare Au NCs, the synthesized Zr(IV)/Au NCs showed stronger orange fluorescence at 610 nm. On the other hand, the Zr(IV)/Au NCs retained the phosphatase-like activity of Zr(IV) ions and could catalyze the hydrolysis of fluorescent substrate 4-methylumbelliferyl phosphate to produce blue emission at 450 nm. The addition of phosphate salts could effectively inhibit the catalytic activity of Zr(IV)/Au NCs, resulting the fluorescence decrease at 450 nm. However, the fluorescence at 610 nm almost unchanged upon the addition of phosphates. Based on this finding, the ratiometric detection of phosphates using the fluorescence intensity ratio (I450/I610) was demonstrated. The method has been further applied for sensing total phosphates in frozen shrimp samples with satisfactory results.

Zirconium-amino acid framework as a green phosphatase-like nanozyme for the selective detection of phosphate-containing drugs.

The zirconium-amino acid framework MIP-202(Zr) was reported as a green phosphatase-like nanozyme for the first time. Moreover, its phosphatase-like activity can be inhibited by phosphate-containing drugs. Based on this finding, a universal fluorimetric strategy for sensing phosphate-containing drugs was developed. The detection limit was as low as 2 ng mL-1 for the model drug alendronate sodium. This strategy exhibits excellent selectivity over other non-phosphate-containing drugs and will broaden the applications of phosphatase-like nanozymes in clinical pharmacy.

Highly sensitive detection of Tb3+ and ATP based on a novel asymmetric anthracene derivative.

A novel fluorescent sensor based on an asymmetric anthracene derivative (SSAPA) was designed and synthesized. Using this molecule, a rapid and sensitive assay for detecting Tb3+ and ATP in aqueous solutions was established. The SSAPA molecule had excellent aggregation-induced emission (AIE) performance and good aqueous dispersion ability. This molecule could coordinate with Tb3+ and the fluorescence quenched linearly with the increase in the concentration of Tb3+ from 0.005 to 1.2 µM. Since both Tb3+ and adenosine triphosphate (ATP) have strong binding ability, ATP can compete with Tb3+ from the SSAPA/Tb3+ complex leading to fluorescence recovery. In this way, a brand-new fluorescent "turn-on" assay for ATP in the range from 0.01 to 0.4 µM was developed using the Tb3+-based complex probe. The detection limits for Tb3+ and ATP both reached single-digit nanomole per millilitre (2.8 nM and 4.5 nM, respectively), which demonstrated that this method has high sensitivity. Besides, Tb3+ and ATP also could be well detected in other complex environments such as real water samples or serum samples. This study provides a feasible assay for detecting trace amounts of Tb3+ and ATP in aqueous solutions.

Design, synthesis and imaging of a novel mitochondrial fluorescent nanoprobe based on distyreneanthracene-substituted triphenylphosphonium salt.

Targeting and monitoring the dynamics of mitochondria are of great significance because mitochondria are involved in a variety of physiological and pathological processes. For achieving this purpose, highly sensitive, photostable, tolerance and specific fluorescent probe is necessary. To obtain a superior mitochondrial fluorescent probe, (4-distyreneanthracenoxybutyl) bis(triphenylphosphonium) bromide (DSA-TPP) with aggregation-induced emission (AIE) characteristic was designed and synthesized for mitochondrial targeting. DSA-TPP dots with high fluorescence quantum yield (Φ = 17.9) and small particle size (8 nm) can be easily prepared by self-assembly formation. DSA-TPP dots had the ability of lightning mitochondria in living cells with high brightness, superior photostability and strong tolerance to cell environment change.

Chemiluminescence of the Ce(IV)/CDP-Star System Based on the Phosphatase-like Activity of Ce(IV) Ions.

The phosphatase-like activity of Ce(IV) ions was applied for chemiluminescence (CL) analysis for the first time. Ce(IV) can catalyze the hydrolysis of CDP-star, which is a phosphatase substrate, to produce strong CL emission. The CL performance of the Ce(IV)/CDP-star system can be significantly improved by the addition of ionic liquids. In the presence of 1-butyl-3-methylimidazolium tetrafluoroborate, the selective and sensitive CL detection of Ce(IV) ions was achieved with a detection limit of 460 nM. The proposed CL system was also used for the detection of ascorbic acid and ClO-. It is based on the phenomenon that Ce(IV) can catalyze the hydrolysis of CDP-star, while Ce(III) cannot. The introduction of reductive ascorbic acid into the mixture of Ce(IV)/CDP-star can turn off the CL signal, while the addition of oxidative ClO- into the solution of Ce(III)/CDP-star can turn on the CL emission. Finally, Ce(IV)/CDP-star CL was successfully applied for evaluating the total antioxidant capacity in commercial fruit juice samples.

Ionic liquid-assisted chemiluminescent immunoassay of prostate specific antigen using nanoceria as an alkaline phosphatase-like nanozyme label.

An alkaline phosphatase-like nanozyme was applied for an immunoassay for the first time. By using nanoceria as the alkaline phosphatase-like catalytic label and CDP-star as the substrate, the chemiluminescent detection of prostate specific antigen was demonstrated. More importantly, the addition of ionic liquid can significantly increase the sensitivity of the immunoassay. With the aid of ionic liquid, an order of magnitude improvement in the sensitivity was achieved with a detection limit of 53 fg mL-1.

Right-/left-handed helical G-quartet nanostructures with full-color and energy transfer circularly polarized luminescence.

Right (R)- and left (L)-handed helical G-quartet nanostructures were synthesized for the first time simultaneously via the self-assembly of 5'-guanosine monophosphate (GMP), the helical handedness of which is well regulated by metal ions. These g-nanostructures were further applied as circularly polarized luminescence (CPL) templates to realize full-color R-/L-CPL and Förster resonance energy transfer CPL. The glum value reached 10-2, indicating their excellent template function for CPL materials design and application.

Fluorescent aptasensors for parallel analysis of biomolecules based on interlocked DNA catenane nanomachines.

Stimuli-responsive DNA catenane nanomachines have received considerable interest in the area of DNA nanotechnology. However, the sensing and bio-sensing applications of DNA catenane nanomachines are rarely explored. Herein, the biological small molecule and protein responsive DNA catenane nanomachines were designed by utilizing the specific aptamer/target interaction. In the presence of the target, the blocked catalytically inactive DNA catenane can be activated, while generating the metal ion-dependent DNAzyme as the signal output unit. The method allows the fluorescent detection of ATP and lysozyme with a detection limit of 20 nM and 200 pM, respectively. More importantly, through the use of different fluorophores as labels, the parallel analysis of ATP and lysozyme in the mixed solution was demonstrated based on two interlocked DNA catenanes. Compared with linear scaffold, the interlocked DNA rings showed enhanced stability against enzyme degradation and easily realized intramolecular reconfiguration. Due to its inherent advantages, the interlocked DNA catenane holds great promise in the field of DNA based sensors and nanodevices.

The phosphatase-like activity of zirconium oxide nanoparticles and their application in near-infrared intracellular imaging.

In this study, the phosphatase mimetic activity of zirconium oxide nanoparticles (ZrO2 NPs) has been demonstrated. They can effectively catalyze the dephosphorylation of a series of commercial fluorogenic and chromogenic substrates of natural phosphatases. Compared with natural phosphatases, ZrO2 NPs possess several advantages such as low cost, facile preparation procedures, and high stability in a broader pH range or at high temperatures. In addition, the activity of ZrO2 NPs toward some important biomolecules was investigated. The ZrO2 NPs can catalyze the dephosphorylation of ATP and o-phospho-l-tyrosine, but they cannot react with DNA strands. These data are important for the further bio-related applications of ZrO2 NPs. Finally, the potential application of ZrO2 NPs in intracellular imaging is also demonstrated by using a near-infrared fluorescent substrate of alkaline phosphatase.

Highly sensitive chemiluminescent sensing of intracellular Al3+ based on the phosphatase mimetic activity of cerium oxide nanoparticles.

Nanomaterials with enzyme-like characteristics (also called nanozymes) have attracted increasing attention in the area of analytical chemistry. Nevertheless, most of the nanozymes used for analytical applications are oxidoreductase mimics, and their enzyme-like activities are usually demonstrated by using chromogenic and/or fluorogenic substrates. Herein, the phosphatase mimetic activity of cerium oxide nanoparticles (nanoceria) was investigated by using CDP-star as the chemiluminescent (CL) substrate. Interestingly, we found that the phosphatase mimetic activity of nanoceria can be remarkably inhibited by the addition of Al3+. Based on this finding, a highly sensitive and selective CL method for Al3+ detection is proposed. The CL intensity of the nanoceria/CDP-star system decreased with the increasing Al3+ concentrations in the range from 30 nM to 3.5 µM. A detection limit as low as 10 nM was obtained. Finally, the CL detection of intracellular Al3+ was achieved, demonstrating the utility of the CL method in complex biological samples.

Electrochemiluminescence resonance energy transfer immunoassay for alkaline phosphatase using p-nitrophenyl phosphate as substrate.

A sensitive electrochemiluminescent immunoassay for alkaline phosphatase (ALP) using p-nitrophenyl phosphate (PNPP) as substrate based on the electrochemiluminescence resonance energy transfer (ECRET) is developed. Luminol-doped silica nanoparticles (luminol-SiNPs) are prepared by water/oil (W/O) microemulsion method. PNPP convertes to p-nitrophenol (PNP) in the presence of ALP, which results in the absorption peak shifting from 360 nm to 450 nm. Herein the spectral overlap between absorption spectrum of PNP and electrochemiluminescence (ECL) spectrum of luminol-SiNPs (425 nm) makes energy transfer occur from luminol-SiNPs to PNP. In the optimized conditions, a linear relationship was obtained using this ECRET method at the concentration of ALP from 5 to 50 U/L (r = 0.9905) and with the limit of detection (LOD) of 0.8 U/L. This ECRET method exhibits sufficient specificity for ALP over other enzymes such as horseradish peroxidase, trypsin and lysozyme.

Fluorescence detection of protamine, heparin and heparinase II based on a novel AIE molecule with four carboxyl.

In this research, a new DSA (Distyryl-anthracene) derivative with four carboxyl groups was designed and synthesized. This molecule exhibits aggregation-induced emission property (AIE). With the AIE character, a convenient and sensitive fluorescent probe for the detection of protamine, heparin and heparinase has been developed. The protamine can directly induce the aggregation of probe, which caused by electrostatic attraction. In this way, the turn-on detection of protamine is achieved, and the detection limit is as low as 30 ng mL-1. When heparin appears, the probes will be redisperse in solution, which causes a decrease in fluorescence intensity. Besides, this method also shows good selectivity and sensitivity, and the linear range of heparin is from 0.08 to 8 µg mL-1 with detection limit of 37 ng mL-1. After hydrolyzing heparin by heparinase, the probes rebind with protamine and the fluorescence enhance. The fluorescence enhancement was linearly related to the concentration of heparinase in the range of 0.02-2.0 µg mL-1 and detection limit as low as 143.7 ng mL-1. In addition, the results exhibited that the recovery percentage of heparinase in bovine samples reached to 96-101%.

Peroxidase-like Activity of Metal-Organic Framework [Cu(PDA)(DMF)] and Its Application for Colorimetric Detection of Dopamine.

A metal-organic framework (MOF) [Cu(PDA)(DMF)] was synthesized under mild mixed solvothermal conditions. It is constructed by 1,10-phenanthroline-2,9-dicarboxylic acid (H2PDA) and Cu2+ ions. The complex exhibits high peroxidase-like activity and can catalytically oxidize the colorless substrate 3,3',5,5'-tetramethylbenzidine to a blue product in the presence of H2O2. However, the peroxidase-like activity of [Cu(PDA)(DMF)] can be potently inhibited in the presence of dopamine. Based on this phenomenon, the colorimetric detection of dopamine was demonstrated with good selectivity and high sensitivity. [Cu(PDA)(DMF)] showed good stability and robust catalytic activity, which has been employed in the detection of dopamine in human urine and pharmaceutical samples.

A colorimetric heparin assay based on the inhibition of the oxidase mimicking activity of cerium oxide nanoparticles.

A colorimetric method is described for the sensitive detection of heparin (Hep). It is based on the finding that Hep can effectively inhibit the oxidase mimicking activity of cerium oxide nanoparticles (nanoceria). In the presence of Hep, the catalytic activity of nanoceria toward the oxidation of the chromogenic substrate 3,3',5,5'-tetramethylbenzidine by oxygen is strongly decreased. The inhibition mechanism is attributed to the fact that Hep is adsorbed on the surface of the nanoceria. Under optimal condition, the absorbance (measured at 652 nm) decreases with increasing Hep concentrations in the range from 30 to 700 nM. The detection limit is 20 nM. The method was applied to the determination of Hep in medical injection sample and serum sample with satisfactory results. Graphical abstract Schematic presentation of the inhibition of oxidase-like activity of nanoceria by heparin. This allows the sensitive detection of heparin in medical injection sample and serum sample with satisfactory results.