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Protein drugs showing strong pharmaceutical activity, high specificity, and low toxicity and side effects have drawn extensive attention in the field of life sciences and medicine. Precise evaluation of the function of these drugs requires accurate and sensitive detection methods. Here, we report a novel chromatography-tandem mass spectrometry (LC-MS/MS) method for sensitive and selective detection of protein drugs. Magnetic nanoparticles (Apt29@MNPs) were functionalized by thrombin aptamers, and quantum dots (Apt15@ss@QDs) were dual-functionalized with quantitative thrombin aptamers and small molecules with high ionization efficiency as the mass barcode. After Apt29@MNPs specifically purify and enrich thrombin from biological samples, they can form a nano "sandwich structure" when Apt15@ss@QDs are added, resulting in the release of the mass barcode for LC-MS/MS analysis via the cutting of the disulfide bond. Since there is a higher quantitative molecular ratio of mass barcode to thrombin in the nano-"sandwich structure", quantitative detection of thrombin with high sensitivity and selectivity can be achieved via the LC-MS/MS detection of the mass barcode with high ionization efficiency rather than thrombin, which effectively avoids the disadvantages of direct protein detection by mass spectrometry. The established method for thrombin detection shows a good linear relationship in a concentration range of 0.00115-1.15 nM with a limit of detection (LOD) of 0.0007 nM. The present work provides a new approach for the effective and sensitive quantitative analysis of protein drugs and would be of great significance in promoting the development of protein drugs and clinical applications.

A standard desktop printer with multiple ink cartridges can accurately deposit a broad variety of biomaterials on microfluidic paper-based analytical devices (μPADs) which have been extensively applied to environmental monitoring and screening of food and beverage contamination. Finding ways to realize sample quantitative control by tuning the CMYK value, however, remains challenging. Herein, we studied the influence of the CMYK value on the ink volume jetted by ink cartridges. The regularity research on a single-color and two-colors was performed in two print mode-grayscale printing and color printing. The results demonstrated that the number of ink dots increased with the increase of the gray value and opacity value, which means that the amount of the bio-ink increases with the increase of the CMYK value. The 3,3',5,5'-tetramethylbenzidine-horseradish peroxidase-hydrogen peroxide, glucose oxidase-horseradish peroxidase and bull serum albumin-citrate buffer-tetrabromophenol blue systems were chosen as examples to prove the print regularity. Samples and assay reagents can be quantitatively deposited on a substrate by adjusting the CMYK value with as many as four ink cartridges. The present approach has been successfully applied to assay the targets in real serum samples, showing the potential application of the most common office piezoelectric printer in μPADs.

We herein report the intrinsic alkaline peroxidase-like activity exhibited by CuO nanoparticles when 3-(4-hydroxyphenyl)propionic acid was employed as a substrate. Based on this observation, a fluorometric assay method with a low detection limit of 0.81 μM was established for H 2 O 2 determination under alkaline conditions. Notably, ammonia was found to inhibit the alkaline peroxidase-like activity of the CuO nanoparticles. Thus, a sensing platform for the determination of urea and urease was successfully constructed, with the limits of detection for urea and urease being 27 μM and 2.6 U L -1 , respectively. This platform was then applied for the detection of urea in human urine and urease in soil, which yielded satisfactory results. These results suggest that it is possible to extend the catalytic potential of peroxidase and its mimetics from acidic and neutral conditions to include activity in alkaline media as well. Furthermore, this strategy is a novel method for the analysis of urea and urease. The assay developed in this work is facile, inexpensive, convenient, and highly selective and sensitive. Therefore, it is expected that this system can serve as a template for the development of similar enzyme nano-mimics.

Given the importance of hydrogen peroxide (H2O2) in many biological processes and its wide application in various industries, the demand for sensitive, accurate, and economical H2O2 sensors is high. In this study, we used Fenton reaction-stimulated fluorescence quenching of N-acetyl-L-cysteine-protected gold nanoclusters (NAC-AuNCs) as a reporter system for the determination of H2O2. After the experimental conditions were optimized, the sensing platform enabled the analysis of H2O2 with a limit of detection (LOD) as low as 0.027 μM. As the glucose oxidase cascade leads to the generation of H2O2 and catalase catalyzes the decomposition of H2O2, these two biocatalytic procedures can be probed by the Fenton reaction-mediated quenching of NAC-AuNCs. The LOD for glucose was found to be 0.18 μM, and the linear range was 0.39-27.22 μM. The LOD for catalase was 0.002 U mL(-1), and the linear range was 0.01-0.3 U mL(-1). Moreover, the proposed sensing methods were successfully applied for human serum glucose detection and the non-invasive determination of catalase activity in human saliva, demonstrating their great potential for practical applications.

A facile approach is proposed for the growth of platinum nanoparticles on graphene oxide (PtNPs/GO). The resulting PtNPs/GO hybrid has been proved to function as peroxidase mimics that can catalyze the oxidation of peroxidase substrates in the presence of hydrogen peroxide (H2O2). Kinetic studies indicate that the PtNPs/GO nanocomposite has a considerably higher affinity for both 3,3',5,5'-tetramethylbenzidine (TMB) and H2O2 than those of other platinum-based peroxidase mimics. Furthermore, colorimetric recognition and sensing of L-cysteine with high sensitivity and selectivity is presented based on target-induced shielding against the peroxidase-like activity of PtNPs/GO. We envision that this material will be an ideal candidate for a wide range of potential applications in the fields of biomedicine and environmental chemistry.

Hydroxyl radicals (˙OH) play an important role in human diseases. Traditional detection methods are time consuming and require expensive instruments. Here, we present a simple and sensitive method for the detection of hydroxyl radicals on a microfluidic chip using an electrochemical technique. Aniline monomer is electrochemically polymerized on the surface of a pencil graphite electrode and carbonized at 800 °C. The resulting N-doped porous carbon nanofiber-modified pencil graphite electrode is embedded into a microfluidic chip directly as a working electrode. 4-Hydroxybenzoic acid (4-HBA) is selected as the trapping agent owing to its unique 3,4-DHBA product and high trapping efficiency. A low detection limit of 1.0 × 10(-6) M is achieved on the microfluidic chip. As a demonstration, the microfluidic chip is successfully utilized for the detection of ˙OH in cigarette smoke. The strong π-π stacking and hydrophobic interactions between the nitrogen-doped carbon materials and the pencil graphite make the modified electrode well-suited for the microfluidic chip.

Herein we developed an IMPLICATION logic gate based on citrate-capped AuNPs by employing thiocyanate (SCN(-)) and iodide (I(-)) as inputs, and devised a colorimetric sensor for the determination of I(-) with good selectivity and sensitivity. To the best of our knowledge, this is the first example in which two species of anions serve as inputs to obtain visually observed Boolean outputs. Under the optimum conditions, 0.8 μM I(-) could induce a significant color change and be recognized by the naked eye. The detection limit is 50 nM by using UV-vis spectroscopy.

With the advantages of in situ analysis and high surface sensitivity, surface-enhanced infrared absorption spectroscopy in attenuated total reflection mode (ATR-SEIRAS) combined with electrochemical methods has been employed to examine the interfacial direct electron transfer activity and adsorption kinetics of cytochrome c (cyt c). This work presents data on cyt c adsorption onto negatively charged mercaptohexanoic acid (MHA) and positively charged 6-amino-1-hexanethiol (MHN) self-assembled monolayers (SAMs) on gold nanofilm surfaces. The adsorbed cyt c displays a higher apparent electron transfer rate constant (33.5 ± 2.4 s(-1)) and apparent binding rate constant (73.1 ± 5.2 M(-1) s(-1)) at the MHA SAMs surface than those on the MHN SAMs surface. The results demonstrate that the surface charge density determines the protein adsorption kinetics, while the surface charge character determines the conformation and orientation of proteins assembled which in turn affects the direct electron transfer activity.

An electrochemically induced sol-gel process has been used to efficiently immobilize Ru(bpy)(3)(2+) in a 3D porous silica film matrix deposited on a glassy carbon electrode (GCE), forming a solid state electro-chemiluminescence (ECL) sensor. In this approach, electrolysis of the GCE at cathodic voltages from a solution of ammonium fluorosilicate containing Ru(bpy)(3)(2+) results in the reduction of water to hydroxyl ions and hydrogen bubbles. The former product catalyzes the hydrolysis of ammonium fluorosilicate to form a silica film; while the hydrogen bubbles act as a dynamic template in forming a porous silica matrix. Therefore, a large quantity of Ru(bpy)(3)(2+) ions can be efficiently encapsulated in the porous silica matrix, and the formed porous structure offers a good mass transport path. The fabricated [Ru(bpy)(3)](2+) solid-state ECL sensor shows high sensitivity and stability towards the determination of tripropylamine (TPA). The electrochemically generated luminescence signal shows a good linear relationship to TPA concentration ranging from 3.46 × 10(-10) to 3.70 × 10(-6) M and 3.70 × 10(-6) to 3.60 × 10(-4) M, with an extremely low detection limit of 17 pM (S/N = 3). The present approach is effective for encapsulation of various molecules and could find wide application in the construction of various sensors.

In this study, we demonstrate a more sensitive colorimetric assay for the detection of melamine based on the strong attraction between exocyclic amine groups of melamine and surface-bound AuCl(4)(-)/AuCl(2)(-) ions on bare gold nanoparticles prepared by the borohydride reduction method. This method is rather simple without further modification of gold nanoparticles and complicated pretreatment of samples. The detection limit of the method is 2.0 × 10(-7) g L(-1) for melamine in infant formula, which is much lower (about 1000-fold) than that of using citrate-capped GNPs as a colorimetric probe. The proposed scheme could be a good alternative means for on-site and real-time screening of melamine adulterant in infant formula without costly instruments.

Water-soluble cupric oxide nanoparticles are fabricated via a quick-precipitation method and used as peroxidase mimetics for ultrasensitive detection of hydrogen peroxide and glucose. The water-soluble CuO nanoparticles show much higher catalytic activity than that of commercial CuO nanoparticles due to their higher affinity to hydrogen peroxide. In addition, the as-prepared CuO nanoparticles are stable over a wide range of pH and temperature. This excellent stability in the form of aqueous colloidal suspensions makes the application of the water-soluble CuO nanoparticles easier in aqueous systems. A colorimetric assay for hydrogen peroxide and glucose has been established based on the catalytic oxidation of phenol coupled with 4-amino-atipyrine by the action of hydrogen peroxide. This analytical platform not only confirms the intrinsic peroxidase-like activity of the water-soluble cupric oxide nanoparticles, but also shows its great potential applications in environmental chemistry, biotechnology and medicine.