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An electrochemical platform based on a screen-printed carbon electrode (SPCE) is developed to detect parathyroid hormone (PTH). A nanocomposite of multi-walled carbon nanotube (MWCNT) and gold nanoparticles (AuNP) was deposited on the SPCE to immobilize antibodies and horseradish peroxidase (HRP). MWCNT improved the stability and conductivity of the immunosensor because of its good electron-transfer ability and tubular structure. The AuNP not only provided a large surface area for antibody immobilization, but it also enhanced the electrochemical signal for enzyme-linked immunosensing. Cyclic voltammetry showed both electron transfer and the effective surface area were increased on the modified electrode. The characteristics of the modified SPCE were assayed by Raman spectroscopy, scanning electron microscopy, atomic force microscopy, and electrochemical techniques. The linear detection range of this PTH immunosensor was within 1-300 pg/ml, and the electrochemical performance was not affected by interference from protein components in human serum. After storage at 4 °C for 28 days, 85% PTH sensing ability of this immunosensor was maintained compared to the freshly prepared one using the SWV and DPV methods. The relative standard deviations of all measurements were within 3-8% for both voltammetric methods. These results indicated the developed immunosensor had good stability and reproducibility. This PTH immunosensor had a detection limit of 0.886 and 0.065 pg/ml for the differential pulse voltammetry and square wave voltammetry, respectively. We provided a quick analysis of serum PTH which might be used as an electrochemical immunosensing platform for point-of-care testing.Fabrication of non-enzymatic electrochemical sensors based on metal oxides with low valence-state for nanomolar detection of H2O2 has been a great challenge. In this work, a novel neuron-network-like Cu-MoO2/C hierarchical structure was simply prepared by in-situ pyrolysis of 3D bimetallic-organic framework [Cu(Mo2O7)L]n [L N-(pyridin-3-ylmethyl)pyridine-2-amine] crystals. Meanwhile, the MoO2/C nano-aggregates were also obtained by liquid phase copper etching. Subsequently, two non-enzymatic electrochemical sensors were fabricated by simple drop-coating of the above two materials on the surface of glassy carbon electrode (GCE). Electrochemical measurements indicate that the Cu-MoO2/C/GCE possesses highly efficient electrocatalytic H2O2 property during wider linear range of 0.24 μM-3.27 mM. At room temperature, the Cu-MoO2/C composite displays higher sensitivity (233.4 μA mM-1 cm-2) and lower limit of detection (LOD = 85 nM), which are 1 and 2.5 times larger than those of MoO2/C material, respectively. Such excellent ability for trace H2O2 detection mainly originates from the synergism of neuron-network-like structure, enhanced electrical conductivity and increased active sites caused by low valence-state MoO2 and co-doping of Cu and carbon, and even the interaction between Cu and Mo. In addition, the H2O2 detection in spiked human serum and commercially real samples indicates that the Cu-MoO2/C/GCE sensor has certain potential application in the fields of environment and biology.Hydrogen deuterium exchange coupled with mass spectrometry (HDX-MS) is a powerful technique for the characterization of protein dynamics and protein interactions. Recent technological developments in the HDX-MS field, such as sub-zero LC separations, large-scale data analysis tools, and efficient protein digestion methods, have allowed for the application of HDX-MS to the analysis of multi protein systems in addition to pure protein analysis. Still, high-throughput HDX-MS analysis of complex samples is not widespread because the co-elution of peptides combined with increased peak complexity after labeling makes peak de-convolution extremely difficult. Here, for the first time, we evaluated and optimized long gradient subzero-temperature ultra-high-pressure liquid chromatography (UPLC) separation conditions for the HDX-MS analysis of complex protein samples such as E. coli cell lysate digest. Under the optimized conditions, we identified 1419 deuterated peptides from 320 proteins at -10 °C, which is about 3-fold more when compared with a 15-min gradient separation under the same conditions. Interestingly, our results suggested that the peptides eluted late in the gradient are well-protected by peptide-column interactions at -10 °C so that peptides eluted even at the end of the gradient maintain high levels of deuteration. Overall, our study suggests that the optimized, sub-zero, long-gradient UPLC separation is capable of characterizing thousands of peptides in a single HDX-MS analysis with low back-exchange rates. As a result, this technique holds great potential for characterizing complex samples such as cell lysates using HDX-MS.Lactose (LAC) is a disaccharide - major sugar, present in milk and dairy products. LAC content is an important indicator of milk quality and abnormalities in food industries, as well as in human and animal health. The present study reports the development of an innovative imprinted voltammetric sensor for sensitive detection of LAC. check details The sensor was constructed using electropolymerized pyrrole (Py) molecularly imprinted polymer (MIP) on graphite paper electrode (PE). The MIP film was constructed through the electrosynthesis of polypyrrole (PPy) in the presence of LAC (template molecule) on PE (PPy/PE). To optimize the detection conditions, several factors affecting the PPy/PE sensor performance were assessed by multivariate methods (Plackett-Burman design and central composite design). Under optimized conditions, the proposed analytical method was applied for LAC detection in whole and LAC-free milks, where it demonstrated high sensitivity and selectivity, with two dynamic linear ranges of concentration (1.0-10 nmol L-1 and 25-125 nmol L-1) and a detection limit of 0.88 nmol L-1. The MIP sensor showed selective molecular recognition for LAC in the presence of structurally related molecules. The proposed PPy/PE sensor exhibited good stability, as well as excellent reproducibility and repeatability. Based on the results obtained, the PPy/PE is found to be highly promising for sensitive detection of LAC.Copper ions (Cu2+) pollution in the water environment poses a great threat to the health function of life-sustaining metabolic activities. However, the current detection methods need relatively expensive instruments, complex operation procedures and long time, so a facile and direct detection method is desired to be developed. In this work, the Ni-based composite wires with p-n junction (the Ni/NiO/ZnO/Chitosan wire) and Schottky junction (the Ni/NiO/Au/Chitosan wire) were fabricated, and the barrier driven electrochemical sensing mechanism was studied. The direct and facile detection of Cu2+ was achieved with a wide linear range (0-6000 nM) and a low LOD (0.81 nM). The excellent stability and recovery in real water samples made the Ni-based composite wires a promising candidate for the practical application. The interfacial barriers of semiconductor can be used as a special sensing factor to develop novel sensors.Phospholipase A2 (PLA2) may be a vital biomarker for the prediction and diagnosis of some diseases. Consequently, it is of great significance to quantitatively detect PLA2 in biologic samples. Herein, on the basis of the principle of luminescence resonance energy transfer (LRET) between upconversion nanoparticles (UCNPs) and SYBR Green I (SG), we proposed a technology for the highly sensitive detection of PLA2 amount. Therein, as an energy receptor, SG will be quantitatively loaded into liposomes firstly. Then, due to the hydrolysis of liposomes under the catalysis of PLA2, SG will be released and inserted into the double-stranded DNA (dsDNA) on the surface of UCNPs, which triggers the LRET because of the shortening of effective spatial distance between UCNPs and SG. Under exciting of NIR light, UCNPs emit luminescence at 476 nm, which makes SG emit fluorescence at 522 nm through LRET. Under optimal conditions, the emission intensity ratio (I522 nm/I476 nm) increased linearly with the PLA2 amount in the range of 20 U/L to 400 U/L, and the limit of detection (LOD) reached 15 U/L. Here, after comparing with the clinical standard method, it is found that the biosensor is expected to provide a convenient and sensitive assay for the detection of PLA2 in actual serum samples. Furthermore, such biosensor can also be used to test the inhibitor of PLA2.This study demonstrated a facile, green and bioinspired approach to synthesize protein-posnjakite nanobiohybrid with rod-assembled hollow shuttle-like structure. Through the one-pot mild coprecipitation process, the inorganic mineral posnjakite (Cu4(SO4) (OH)6·H2O) served as a nanocarrier to efficient co-immobilization of recognition protein (streptavidin) and enzyme (horseradish peroxidase) for signal amplification, which avoids tedious linking or purification procedures and significantly simplifies the synthetic process. This nanobiohybrid was then utilized as the signal tag for immunoassays and presented excellent performance for the detection of insecticidal crystalline (Cry) protein Cry1Ab, in the linear range of 0.1-40 ng mL-1, with the limit of detection of 63 pg mL-1. This proposed strategy is expected to the integration of a variety of biomolecules with posnjakite to design diverse multifunctional nanobiohybrids for multiple applications extending from biosensors, catalysis and biomedicine to environmental science and energy.A polymer-film inertial microfluidic jigsaw (PIMJ) sorter with trapezoidal spiral channels using the jigsaw puzzle method was proposed to realize precise and high-throughput rare cell separation. The PIMJ sorter was fabricated by assembling laser-patterned polymer-film layers of different thicknesses. After illustrating the conceptual design and fabrication process, the effects of the cross-sectional dimension, particle size, and operational flow rate on particle focusing were systematically explored under a broad flow rate range. Then, the separation performances of the PIMJ sorter were characterized using the binary particle mixture and the blood samples spiked with four types of tumor cells. The results indicated that the complete separation of the binary particles with a minimum size difference of 2 μm was successfully realized at a high throughput up to 3000 μL/min. A high recovery ratio of 90.57%-94.14% and a high purity of 48.67%-79.05% were achieved for the separation of rare tumor cells from white blood cells (WBCs). Finally, the PIMJ sorter was successfully employed for separating rare circulating tumor cells (CTCs) from the metastatic breast and lung cancer patients with a capture ratio of 7-226 CTCs per 5 mL sample. The results proved the high sensitivity and high reliability of the PIMJ sorter. The PIMJ sorter is expected to be a potential device for precise CTC separation towards the clinical applications.Determination of complex pollutants often involves many high-cost and laborious operations. Today's pop machine-learning (ML) technology has exhibited their amazing successes in image recognition, drug designing, disease detection, natural language understanding, etc. ML-driven samples testing will inevitably promote the development of related subjects and fields, but the biggest challenge ahead for this process is how to provide some intelligible and sufficient data for various algorithms. In this work, we present a full strategy for rapid detecting mixed pollutants through the synergistic application of holographic spectrum and convolutional neural network (CNN). The results have shown that a well-trained CNN model could realize quantitative analysis of the mixed pollutants by extracting spectral information of matters, suggesting the strategy's value in facilitating the study of complex chemical systems.
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