Hua-Ping Peng

Water-soluble gold nanoclusters prepared by protein-ligand interaction as fluorescent probe for real-time assay of pyrophosphatase activity.

This paper reports a new and facile method for the synthesis of water-soluble thiolate-protected AuNCs via protein-ligand interaction. Using 3-mercaptopropionic acid (MPA) as a model ligand and bovine serum albumin (BSA) as a model protein, water-soluble AuNCs (BSA/MPA-AuNCs) with intense orange-yellow fluorescent emission (quantum yield=16%) are obtained. Results show that AuNCs produced with this method have hydrophobic interactions with BSA. The synthetic strategy is then successfully extended to produce water-soluble AuNCs protected by other thiolates. Moreover, a sensitive and eco-friendly sensing system is established for detection of the activity of inorganic pyrophosphatase (PPase), which relies on the selective coordination of Fe(3+)with BSA/MPA-AuNCs, the higher affinity between pyrophosphate (PPi) and Fe(3+), and the hydrolysis of PPi by PPase. A good linearity between the fluorescence intensity and PPase activity within the range from 0.1 to 3U/L is found, with a detection limit down to 0.07U/L. Additionally, the fluorescent assay developed here is utilized to assay the PPase activity in real biological samples and as well as to evaluate PPase inhibitor, illustrating the great potential for biological analysis.

Self-cascade reaction catalyzed by CuO nanoparticle-based dual-functional enzyme mimics

It is desirable but challenging to assemble various biomimetic properties into a functional catalytic cascade system. In this work, cupric oxide nanoparticles were investigated as oxidase mimics for the aerobic oxidation of cysteine to cystine with the generation of hydrogen peroxide. Coupling this property with the peroxidase-like activity of CuO nanoparticles, we constructed a self-organized cascade reaction system based on a single-component nanozyme, which includes the oxidation of cysteine to yield cystine and hydrogen peroxide and the hydrogen peroxide-mediated oxidation of terephthalic acid to produce a fluorescence change. Based on this artificial enzymatic cascade reaction system, a fluorometric assay method with a low detection limit of 6.6nM was established for cysteine determination. This platform was then applied for the detection of cysteine in pharmaceutical products and human plasma, which yielded satisfactory results. Our investigations open up a new route and holds promise for the development and applications of multifunctional nanomaterials as enzyme mimics.

Partially reduced graphene oxide as highly efficient DNA nanoprobe

This work investigates the effect of reduction degree on graphene oxide (GO)-DNA interaction and the fluorescence quenching mechanism. Partial reduced graphene oxide (pRGO), which maintains well water-dispersibility, is synthesized using a mild reduction method by incubating GO suspension under alkaline condition at room temperature. The fluorescence quenching enhances with the restoration degree of sp(2) carbon bonds and follows the static quenching mechanism. The binding constant values imply that pRGO has much stronger affinity with ssDNA than GO. Utilizing this highly efficient nanoprobe, a universal sensing strategy is proposed for homogeneous detection of DNA. Compared with the reported GO-based DNA, this present strategy has obvious advantages such as requirement of low nanoprobe dosage, significantly reduced background, fast fluorescence quenching, and improved sensitivity. Even without any amplification process, the limit of detection can reach as low as 50 pM.

Facile electrochemiluminescence sensing platform based on high-quantum-yield gold nanocluster probe for ultrasensitive glutathione detection

This report outlines a highly sensitive and facile electrochemiluminescence (ECL) sensing platform based on a novel high-quantum-yield Au-nanocluster (AuNC) probe for glutathione (GSH) detection. Owing to the prominent quenching effect of GSH on the ECL of the AuNCs, the proposed ECL nanosensor showed a wide response to GSH in the ranges of 1.0 × 10-9-1.0 × 10-5M and 1.0 × 10-5-1.0 × 10-1M and a low detection limit of 3.2 × 10-10M. In addition, the proposed system exhibited good selectivity for GSH in the presence of other chemical/biological interferences. Moreover, since no further functionalization of AuNC-based sensor interface was necessary, together with the stability, high sensitivity and selectivity of the proposed nanosensor, this convenient approach was able to successfully detect GSH in both of human urine samples and blood samples with excellent recoveries, which indicated its promising application under physiological conditions. Of significant importance is that this study not only helps in gaining a better understanding of the applicability of the ECL properties of AuNCs, but also provides a new avenue for the design and development of ECL sensors based on the novel high-quantum-yield AuNC-based probe and other functional-metal-based NC probes.

Valence States Effect on Electrogenerated Chemiluminescence of Gold Nanocluster

This work elucidated the valence states effect on the electrogenerated chemiluminescence (ECL) performance of gold nanocluster (AuNC). The N-acetyl-l-cysteine-AuNCs (NAC-AuNCs) and the electrochemical reduction method for reducing the AuNCs were first employed to this study. Results demonstrate that the electrochemical reduction degree of the AuNCs depended on the reduction potential, and the enhancement of the ECL signals was positively correlated with the reduction degree of AuNCs, which indicated that the valence state of Au plays a vital role in the ECL performance of AuNCs. Furthermore, the proposed method has been successfully extended to the chemical reduction technique and other nanoclusters. Therefore, an excellent AuNC-based ECL method with various advantages, such as simple preparation, lower toxicity, high sensitivity, and ΦECL, and excellent stability, has been proposed. This approach not only opens up a new avenue for designing and developing ECL device from other functional-metal based NCs, but also extends the huge potential application in the ECL sensing.

Sensitive electrochemical immunosensor based on three-dimensional nanostructure gold electrode.

A sensitive electrochemical immunosensor was developed for detection of alpha-fetoprotein (AFP) based on a three-dimensional nanostructure gold electrode using a facile, rapid, “green” square-wave oxidation-reduction cycle technique. The resulting three-dimensional gold nanocomposites were characterized by scanning electron microscopy and cyclic voltammetry. A “sandwich-type” detection strategy using an electrochemical immunosensor was employed. Under optimal conditions, a good linear relationship between the current response signal and the AFP concentrations was observed in the range of 10–50 ng/mL with a detection limit of 3 pg/mL. This new immunosensor showed a fast amperometric response and high sensitivity and selectivity. It was successfully used to determine AFP in a human serum sample with a relative standard deviation of <5% (n=5). The proposed immunosensor represents a significant step toward practical application in clinical diagnosis and monitoring of prognosis.

An ammonia-based etchant for attaining copper nanoclusters with green fluorescence emission

Luminescent copper nanoclusters (CuNCs) constitute a very active research topic due to their unique properties and lower cost than gold and silver NCs. In this study, we report a new, facile, and rapid top-down etching method for synthesizing luminescent CuNCs, using Cu nanoparticles (CuNPs) as the precursor and ammonia (NH3) as the etchant. The etching mechanism is systematically investigated and the optical and structural properties of the obtained CuNCs are carefully studied. The NH3-triggered etching process is very fast and the newly generated CuNCs can emit strong green fluorescence with a high quantum yield. Moreover, by coupling the urease-catalyzed hydrolysis of urea with the NH3-induced etching of CuNPs, we developed a novel fluorescence turn-on assay for urea. The linear range for urea detection is from 0.25 to 5 mM, and the limit of detection is 0.01 mM. This novel sensing approach, with good sensitivity and excellent selectivity, is then successfully utilized to detect urea in human serum samples, demonstrating its great potential in clinical diagnosis. In addition, the proposed coupling method can be extended to monitor other analytes that influence the size-focusing etching process, allowing metal NCs to be used to construct diverse chemosensors and biosensors.

Gold Nanoparticle-Based Photoluminescent Nanoswitch Controlled by Host–Guest Recognition and Enzymatic Hydrolysis for Arginase Activity Assay

The development of simple yet powerful methods for monitoring enzyme activity is of great significance. Herein, a facile, convenient, cost-effective, and continuous fluorescent method for the detection of arginase and its inhibitor has been reported based on a host-guest interaction-controlled and enzymatic hydrolysis-controlled luminescent nanoswitch. The fluorescence intensity of 6-aza-2-thiothymine-stabilized gold nanoparticle (ATT-AuNP) is enhanced by l-arginine, owing to the formation of a supramolecular host-guest assembly between the guanidine group of l-arginine and ATT molecules capped on the AuNP surface. However, hydrolysis of l-arginine, catalyzed by arginase, leads to a decrease in the fluorescence intensity of l-arginine/ATT-AuNPs hybrids. Upon incorporation of the arginase inhibitor l-norvaline, the fluorescence of the ATT-AuNP-based detecting system is restored. The linear range of arginase activity determination is from 0.0625 to 1.15 U/mL and the limit of detection is 0.056 U/mL. The half-maximal inhibition value IC50 of l-norvaline is determined to be 5.6 mM. The practicability of this luminescent nanoswitch is validated by assaying the arginase activity in rat liver and monitoring the response of rat liver arginase to pharmacological agent. Compared to the existing fluorescent method of arginase activity assay, the approach demonstrated here does not involve any complicated technical manipulation, thereby greatly simplifying the detection steps. We propose that this AuNP-based luminescent nanoswitch would find wide applications in the field of life sciences and medicine.

Label-free, resettable, and multi-readout logic gates based on chemically induced fluorescence switching of gold nanoclusters

Herein, we present a label-free, resettable, multi-readout, and fluorescent logic system by employing bovine serum albumin (BSA) and 3-mercaptopropionic (MPA) co-stabilized gold nanoclusters (BSA/MPA–AuNCs) as a basic work unit. Small chemical molecules including Fe2+, Fe3+, H2O2 and H+/OH− are selected as the inputs to induce the change in fluorescence intensity of BSA/MPA–AuNCs through a non-radiative electron transfer mechanism. Using this novel strategy, a versatile logic system capable of performing elementary logic operations (NAND, NOR, and IMPLICATION) and integrative logic operations (NAND + NOR) can be successfully constructed. More importantly, the logic operations can be also performed on a solid surface in which BSA/MPA–AuNCs are complexed with polyelectrolytes and are coated on a glass slide. We envision that the current work will bring new ideas in establishing chemical logic gates in both liquid media and solid forms.

Electrochemical DNA biosensor based on grafting-to mode of terminal deoxynucleoside transferase-mediated extension

Previously reported electrochemical DNA biosensors based on in-situ polymerization approach reveal that terminal deoxynucleoside transferase (TdTase) has good amplifying performance and promising application in the design of electrochemical DNA biosensor. However, this method, in which the background is significantly affected by the amount of TdTase, suffers from being easy to produce false positive result and poor stability. Herein, we firstly present a novel electrochemical DNA biosensor based on grafting-to mode of TdTase-mediated extension, in which DNA targets are polymerized in homogeneous solution and then hybridized with DNA probes on BSA-based DNA carrier platform. It is surprising to find that the background in the grafting-to mode of TdTase-based electrochemical DNA biosensor have little interference from the employed TdTase. Most importantly, the proposed electrochemical DNA biosensor shows greatly improved detection performance over the in-situ polymerization approach-based electrochemical DNA biosensor.