Distinguishable Magnetic Reporter Coordination with Buoyancy-Magnetism Separation for Immobilization-Free Dual-Target Electrochemical Immunosensing.

Simultaneous sensitive and precise determination of multibiomarkers is of great significance for improving detection efficiency, reducing diagnosis and treatment expenses, and elevating survival rates. However, the development of simple and portable biosensors for simultaneous determination of multiplexed targets in biological fluids still faces challenges. Herein, a unique and versatile immobilization-free dual-target electrochemical biosensing platform, which combines distinguishable magnetic signal reporters with buoyancy-magnetism separation, was designed and constructed for simultaneous detection of carcinoembryonic (CEA) and α-fetoprotein (AFP) in intricate biological fluids. To construct such distinguishable magnetic signal reporters with signal transduction, amplification, and output, secondary antibodies of CEA and AFP were respectively functionalized on methylene blue (MB) and 6-(ferrocenyl)hexanethiol (FeC) modified Fe3O4@Au magnetic nanocomposites. Meanwhile, a multifunctional flotation probe with dual target recognition, capture, and isolation capability was prepared by conjugating primary antibodies (Ab1-CEA, Ab1-AFP) to hollow buoyant microspheres. The target antigens of CEA and AFP can trigger a flotation-mediated sandwich-type immunoreaction and capture a certain amount of the distinguishable magnetic signal reporter, which enables the conversion of the target CEA and AFP quantities to the signal of the potential-resolved MB and FeC. Thus, the MB and FeC currents of magnetically adsorbed distinguishable magnetic reporters can be used to determine the CEA and AFP targets simultaneously and precisely. Accordingly, the proposed strategy exhibited a delightful linear response for CEA and AFP in the range of 100 fg·mL-1-100 ng·mL-1 with detection limits of 33.34 and 17.02 fg·mL-1 (S/N = 3), respectively. Meanwhile, no significant nonspecific adsorption and cross-talk were observed. The biosensing platform has shown satisfactory performance in the determination of real clinical samples. More importantly, the proposed approach can be conveniently extended to universal detection just by simply substituting biorecognition events. Thus, this work opens up a new promising perspective for dual and even multiple targets and offers promising potential applications in clinical diagnosis.

Dual-potential ratiometric electrochemiluminescence based on single emitter and single coreactant for the sensitive detection of carcinoembryonic antigen.

Dual-potential ratiometric electrochemiluminescence (ECL) is in favor of resistance to environmental interference. However, two kinds of emitters or coreactants, and a wide scan potential range (>2 V) are mandatory. This work developed a new dual-potential ratiometric ECL sensor for detection of carcinoembryonic antigen (CEA) using single emitter (luminol) and single coreactant (H2O2) with a mild potential range from -0.1 to 0.6 V. Luminol could produce a strong cathodic ECL (Ec) induced by hydroxyl radicals (HO‧) from the reduction of H2O2, and a relatively weak anodic ECL (Ea). After the ferrocene modified CEA aptamer (Apt-Fc) was attached, Fc could promote Ea by catalyzing the oxidation of H2O2, and reduce Ec by consuming HO‧. With the cycling amplification of the exonuclease I, CEA could substantially reduce the amount of Apt-Fc, resulting in the decrease of Ea and the rise of Ec. So, the ratio of Ec to Ea (Ec/Ea) was used as the detection signal, realizing the sensitive determination of CEA from 0.1 pg mL-1 to 10 ng mL-1 with a LOD of 41.85 fg mL-1 (S/N = 3). The developed sensor demonstrated excellent specificity, stability and reproducibility, with satisfactory results in practical detection.

A perylene diimide electrochemical probe with persulfate as a signal enhancer for dopamine sensing.

Dopamine (DA) is an important biomarker related to parkinsonism, schizophrenia and renal disease. Traditional electrochemical sensors for DA were based on the direct electrochemical oxidation of DA. In this paper, we report a new sensing strategy using N,N'-di(trimethylaminoethyl)perylene diimide (TMPDI) as an electrochemical probe and K2S2O8 as a signal enhancer for DA detection between 0 and -0.7 V with the DPV technique. MoS2 nanoflowers prepared by the hydrothermal method were used as a nanocarrier to load TMPDI. The reduction current of TMPDI was found to show a stepwise and significant increase at -0.24 V with the increase of concentration of K2S2O8 due to the continuous cycle of TMPDI molecules' electrochemical reduction and chemical oxidation. The presence of DA caused a large decrease of the reduction current of TMPDI due to the synergistic interaction of the competitive consumption of DA for K2S2O8 and the blocking effect of polyDA adhering to the electrode surface. The decreased current exhibited a linear response for DA from 10 pM to 100 µM with a detection limit of 4.1 pM and the proposed sensor showed high selectivity and excellent feasibility in human urine/serum sample detection.

A smartphone-based electrochemical POCT for CEA based on signal amplification of Zr6MOFs.

Carcinoembryonic antigen (CEA) is a biomarker of high expression in cancer cells. Highly sensitive and selective detection of CEA holds significant clinical value in the diagnosis, monitoring and efficacy evaluation of malignant tumors. In this work, a smartphone-based electrochemical point-of-care testing (POCT) platform for the detection of CEA was developed based on a Zr6MOF signal amplification strategy. Ferrocene labeled DNA strands (Fc-DNA) were immobilized on Zr6MOFs to form a Fc-DNA/Zr6MOF signal probe. Double-stranded DNA (dsDNA) formed by complementary DNA (cDNA) and CEA aptamer was assembled on a screen-printed electrode via an Au-S bond. When CEA was added, the aptamer specifically bound with CEA, resulting in the exposure of cDNA. Then, Fc-DNA/Zr6MOF signal probes were introduced on the electrode surface through hybridization between Fc-DNA and cDNA. The detection of CEA was realized by measuring the electrochemical response of Fc. The POCT device was made by connecting a modified electrode with a smartphone through a Sensit Smart USB flash disk. Due to the signal amplification of Zr6MOFs, this POCT platform exhibited high sensitivity, wide linear range, and low detection limit for CEA detection. The developed POCT platform has been used for the detection of CEA in actual human serum samples with satisfactory results.

Tungsten-based polyoxometalate nanoclusters with remarkable reactive oxygen species-scavenging activity efficiently quenched luminol-based electrochemiluminescence for sensitive detection of Her-2.

This study aimed to develop a quenching-type electrochemiluminescence (ECL) immunosensor for human epidermal growth factor receptor (Her-2) detection. Firstly, Pd/NiFeOx nanoflowers decorated by in situ formation of gold nanoparticles (Au NPs) and 2D Ti3C2 MXene nanosheets were synthesized (AuPd/NiFeOx/Ti3C2) as carriers to load luminol and primary antibodies. Impressively, AuPd/NiFeOx/Ti3C2 with excellent peroxidase-like activity could accelerate the decomposition of the coreactant H2O2 generating more reactive oxygen species (ROSs) under the working potential from 0 to 0.8 V, resulting in highly efficient ECL emission at 435-nm wavelengths. The introduction of tungsten-based polyoxometalate nanoclusters (W-POM NCs) which exhibit remarkable ROSs-scavenging activity as secondary antibody labels could improve the sensitivity of immunosensors. The ZnO nanoflowers were employed to encapsulate minute-sized W-POM NCs, and polydopamine was self-polymerized on the surface of Zn(W-POM)O to anchor secondary antibodies. The mechanism of the quenching strategy was explored and it was found that W-POM NCs could consume ROSs by the redox reaction of W5+ resulting in W6+. The proposed ECL immunosensor displayed a wide linear response range of 0.1 pg·mL-1 to 50 ng·mL-1, and a low detection limit of 0.036 pg mL-1 (S/N = 3). The recoveries ranged from 93.9 to 99.4%, and the relative standard deviation (RSD) was lower than 10%. This finding is promising for the design of detecting new protein biomarkers.

Tetrahedral DNA nanostructure-corbelled click chemistry-based large-scale assembly of nanozymes for ratiometric fluorescence assay of DNA methyltransferase activity.

Ligation efficiency in a surface-based DNA click chemistry (CuAAC) reaction is extremely restricted by the orientation and density of probes arranged on a heterogeneous surface. Herein, we engineer DNA tetrahedral nanostructure (DTN)-corbelled click chemistry to trigger a hybridization chain reaction (HCR) assembling a large-scale of nanozymes for ratiometric fluorescence detection of DNA adenine methyltransferase (Dam). In this study, a DNA tetrahedron structure with an alkynyl modifying pendant DNA probe (Alk-DTN) is designed and assembled on a magnetic bead (MB) as a scaffold for click chemistry. When a CuO NP-encoded magnetic nanoparticle (CuO-MNP) substrate was methylated by Dam, CuO NPs were released and turned into a mass of Cu+. The Cu+ droves azido modifying lDNA (azide-lDNA) to connect with the Alk-DTN probe on the MB through the click reaction, forming an intact primer to initiate the HCR. The HCR product, a rigid structure double-stranded DNA, periodically assembles glucose oxidase mimicking gold nanoparticles (GNPs) into a large-scale of nanozymes for catalyzing the oxidation of glucose to H2O2. NH2-MIL-101 MOFs, a fluorescent indicator and a biomimetic catalyst, activated the product H2O2 to oxidize o-phenylenediamine (oPD) into visually detectable 2,3-diaminophenazine (DAP). The change of the signal ratio between DAP and NH2-MIL-101 is proportional to the methylation event corresponding to the MTase activity. In this study, the DTN enhances the efficiency of the surface-based DNA click reaction and maintains the catalytic activities of gold nanoparticle nanozymes due to the intrinsic nature of mechanical rigidity and well-controlled orientation and well-adjusted size. Large-scale assembly of nanozymes circumvents the loss of natural enzyme activity caused by chemical modification and greatly improves the amplification efficiency. The proposed biosensor displayed a low detection limit of 0.001 U mL-1 for Dam MTase due to multiple amplification and was effective in real samples and methylation inhibitor screening, providing a promising modular platform for bioanalysis.

An electrochemical ratiometric biosensor for the detection of dopamine based on an MXene-Au nanocomposite.

Compared with single signal detection, a ratiometric biosensor could offer more accurate and reliable results. Here, a ratiometric electrochemical biosensor for the sensitive and accurate detection of dopamine was developed based on the strong adsorption ability of MXene-Au toward methylene blue, an inner reference element. This ratiometric sensing strategy opened up a new avenue for the development of a ratiometric platform.

A label-free electrochemical aptasensor based on Bi-Sb alloy materials for potential POCT of HER-2.

As a prognostic biomarker for breast cancer, human epidermal growth factor receptor 2 (HER-2) is of crucial diagnostic value. Here, a label-free electrochemical aptasensor was established for the ultrasensitive detection of HER-2 using a modified electrode of Bi-Sb alloy materials (Bi-Sb AMs). The performance of the aptasensor was enhanced greatly due to the introduction of Bi-Sb alloy materials (Bi-Sb AMs) with high conductivity. Furthermore, by integrating the aptasensor with the Sensit Smart U-disk electrochemical analyzer, the point-of-care testing (POCT) for HER-2 was realized. Under the optimal experimental parameters, the POCT analyzer showed a wide linear response from 0.01 pg mL-1 to 100 ng mL-1, with a low detection limit (LOD) of 5.96 fg mL-1 for the detection of HER-2. The presented POCT analyzer exhibited good specificity, stability, and reproducibility. Benefiting from the simple operation and rapid testing, the developed analyzer will have potential application in the prognostic diagnosis and treatment of breast cancer.

Fascinating Immobilization-Free Electrochemical Immunosensing Strategy Based on the Cooperation of Buoyancy and Magnetism.

Rapid and accurate detection of biomolecules is of vital importance for the diagnosis of disease and for performing timely treatments. The point-of-care analysis of cancer biomarkers in the blood with low cost and easy processing is still challenging. Herein, an advanced and robust strategy, which integrates the buoyant recognition probe with the magnetic reporter probe in one solution, was first proposed for immobilization-free electrochemical immunosensing. The tumor marker of alpha fetoprotein (AFP) can be captured immune-buoyantly, and then a multifunctional magnetic reporter probe in pseudo-homogeneous solution was further captured to fulfill a sandwich-type immunoreaction. The residual magnetic reporter probe can be firmly and efficiently attracted on a magnetic glassy carbon electrode to fulfill the conversion of the target AFP amount into the residual magnetic electrochemical signal indicator. As a result, the electrochemical signal of methylene blue can accurately reflect the original level of target antigen AFP concentration. By integrating buoyancy-driven quasi-homogenous biorecognition with magnetism-mediated amplification and signal output, the proposed immobilization-free electrochemical immunosensing strategy displayed a wide range of linear response (100 fg mL-1 to 10 ng mL-1), low detection limit (14.52 fg mL-1), and good reproducibility, selectivity, and stability. The designed strategy manifests remarkable advantages including assay simplicity, rapidness, and high sensitivity owing to the in-solution instead of on-electrode biorecognition that could accelerate and improve the biorecognition efficiency. To the best of our knowledge, this is the first cooperation of buoyancy-driven biorecognition with magnetism-mediated signal output in bioanalysis, which would be attractive for rapid clinic biomedical application. Thus, this work provides a fresh perspective for convenient and favorable immobilization-free electrochemical biosensing of universal biomolecules.

Low-temperature molten-salt synthesis of Co3O4 nanoparticles grown on MXene can rapidly remove ornidazole via peroxymonosulfate activation.

We further developed previous work on MXene materials prepared using molten salt methodology. We substituted single, with mixed salts, and reduced the melting point from >724 °C to <360 °C. Cobalt (Co) compounds were simultaneously etched and doped while the MXene material was created using various techniques in which Co compounds occur as Co3O4. The synthesized Co3O4/MXene compound was used as a peroxymonosulfate (PMS) activator that would generate free radicals to degrade antibiotic ornidazole (ONZ). Under optimal conditions, almost 100% of ONZ (30 mg/L) was degraded within 10 min. The Co3O4/MXene + PMS system efficiently degraded ONZ in natural water bodies, and had a broad pH adaptation range (4-11), and strong anion anti-interference. We investigated how the four active substances were generated using radical quenching and electron paramagnetic resonance (EPR) spectroscopy. We identified 12 ONZ intermediates by liquid chromatography-mass spectrometry and propose a plausible degradative mechanism.

Perylene derivative and persulfate as highly efficient electrochemical system for constructing sensitive amperometric aptasensor.

To design highly efficient electrochemistry system was important for construct simple and sensitive biosensors, which was crucial in clinical diagnosis and therapy. In this work, a novel electrochemistry probe N,N'-di (1-hydroxyethyl dimethylaminoethyl) perylene diimide (HDPDI) with positive charges was reported to show two-electron redox behavior in neutral phosphate buffer solution between 0 and -1.0 V. And K2S2O8 in solution could significantly increase the reduction current of HDPDI at -0.29 V, which was interpreted with cyclic catalysis mechanism of K2S2O8. Moreover, HDPDI as electrochemical probe and K2S2O8 as signal enhancer was used to design aptasensors for protein detection. Thrombin was used as target model protein. Thiolate ssDNA with thrombin-binding sequence was immobilized on gold electrode to selectively capture thrombin and adsorb HDPDI. The thiolate ssDNA without binding with thrombin was with random coil structure and could adsorb HDPDI through electrostatic attraction interaction. However, the thiolate ssDNA binding with thrombin became G-quadruplex structure and hardly adsorbed HDPDI. Thus, with increasing the concentration of thrombin, the current signal stepwisely decreased and was taken as detection signal. Compared with other aptasensors based on electrochemistry molecules without signal enhancer, the proposed aptasensors exhibited wider linear response for thrombin between 1 pg mL-1 and 100 ng mL-1 with lower detection limit 0.13 pg mL-1. In addition, the proposed aptasensor showed good feasibility in human serum samples.

Electrochemiluminescence detection of miRNA-21 based on dual signal amplification strategies: Duplex-specific nuclease -mediated target recycle and nicking endonuclease-driven 3D DNA nanomachine.

DNA nanomachines have shown potential application in the construction of various biosensors. Here, an electrochemiluminescence biosensor for the sensitive detection of miRNA-21 were reported based on three-dimensional (3D) DNA nanomachine and duplex-specific nuclease (DSN)-mediated target recycle amplification strategy. First, the bipedal DNA walkers were obtained by DSN-mediated digestion reaction initiated by target miRNA-21.3D DNA tracks were prepared by modifying Fe3O4 magnetic beads (MBs) with ferrocene-labeled DNA (Fc-DNA). The produced DNA walkers autonomously moved along 3D DNA tracks powered by nicking endonuclease. During the movement, ferrocene-labeled DNA was cleaved, resulting in large amounts of Fc-labeled DNA fragments away from the MBs surface. Finally, the liberated Fc-labeled DNA fragments were dropped on the C-g-C3N4 modified electrode surface, leading to the quenching of C-g-C3N4 electrochemiluminescence (ECL). Benefiting from the dual amplification strategy of 3D DNA nanomachine and DSN-mediated target recycling, the developed ECL biosensor exhibited an excellent performance for miRNA-21 detection with a wide linear range of 10 fM to 10 nM and a low detection limit of 1.0 fM. This work offers a new thought for the application of DNA walkers in the construction of various biosensors.

Highly Sensitive Adsorption and Detection of Iodide in Aqueous Solution by a Post-Synthesized Zirconium-Organic Framework.

Effective methods of detection and removal of iodide ions (I-) from radioactive wastewater are urgently needed and developing them remains a great challenge. In this work, an Ag+ decorated stable nano-MOF UiO-66-(COOH)2 was developed for the I- to simultaneously capture and sense in aqueous solution. Due to the uncoordinated carboxylate groups on the UiO-66-(COOH)2 framework, Ag+ was successfully incorporated into the MOF and enhanced the intrinsic fluorescence of MOF. After adding iodide ions, Ag+ would be produced, following the formation of AgI. As a result, Ag+@UiO-66-(COOH)2 can be utilized for the removal of I- in aqueous solution, even in the presence of other common ionic ions (NO2-, NO3-, F-, SO42-). The removal capacity as high as 235.5 mg/g was calculated by Langmuir model; moreover, the fluorescence of Ag+@UiO-66-(COOH)2 gradually decreases with the deposition of AgI, which can be quantitatively depicted by a linear equation. The limit of detection toward I- is calculated to be 0.58 ppm.

Ratiometric Electrochemiluminescence Sensing of Carcinoembryonic Antigen Based on Luminol.

Ratiometric electrochemiluminescence (ECL) sensors can efficiently remove environmental interference to attain precise detection. Nonetheless, two eligible luminophores or coreactants were usually needed, increasing the complexity and restricting their practical application. In this study, a single luminophore of luminol with a single coreactant of H2O2 was employed to construct a dual-potential ratiometric ECL sensor for the detection of carcinoembryonic antigen (CEA). The produced palladium nanoclusters (Pd NCs) employing a DNA duplex as a template could not only stimulate luminol to produce cathodic ECL (Icathodic) but also quench its anodic ECL (Ianodic). During the detection process, CEA could damage the double-stranded structure and reduce the Pd NCs' amount, triggering a significant decrease in the ratio of Icathodic to Ianodic (Icathodic/Ianodic) and thereby achieving sensitive CEA's detection. Furthermore, the Icathodic/Ianodic was independent of the H2O2 concentration, which avoided a prejudicial effect from H2O2 decomposition and considerably enhanced the detection's reliability. The developed ratiometric ECL sensor demonstrated a sensitive detection toward CEA with a wide linear range from 100 ag/mL to 10 ng/mL and a detection limit of 87.1 ag/mL (S/N = 3). In conclusion, this study offers a new idea for constructing ratiometric ECL sensors based on a single luminophore and technical support for cancer's early diagnosis.

Latissimus Dorsi Muscle Flap for Scalp Reconstruction and Postoperative Ulceration Management.

ABSTRACT: The latissimus dorsi muscle (LDM) flap has been widely accepted as the best choice for subtotal or total scalp reconstruction. Because of the unique anatomic and functional features of scalp, ulcerations formation would occur after reconstructive surgeries. in this study, we are presenting a patient with a large scalp defect successfully reconstructed by a latissimus dorsi muscle free flap. Ulcerations with skull exposure formed on the transplanted flap after the first surgery. They were subsequently repaired by flap recycling and tissue expansion techniques. An excellent reconstructive outcome was achieved at the 30-month follow-up after the last surgery and no further complication was found. This clinical report highlights the possibility of ulcer formation after scalp reconstructive surgeries and supports the use of recycle flaps and tissue expanders to manage postoperative ulcerations after latissimus dorsi muscle free flap transplantation.

A signal-amplification electrochemiluminescence sensor based on layer-by-layer assembly of perylene diimide derivatives for dopamine detection at low potential.

Perylene diimide derivatives (PDIs) are suitable ECL luminophore candidates with low triggering potentials and strong ECL signals for fundamental studies and practical applications. However, PDIs tend to aggregate, which affects their optical properties and limits their application in bio-imaging and bio-sensing fields. In this study, an ECL sensor is fabricated based on the layer-by-layer (LBL) assembly of N, N-bis(phosphonomethyl)-3,4,9,10-perylene diimide (PMPDI) and ZrIV ions on the surface of a mesoporous indium tin oxide (ITO) substrate. When six layers of PMPDI are immobilized on ITO, the resulting PMPDI6/ITO electrode shows maximum ECL intensity with K2S2O8 as a co-reactant in the potential range 0 to -0.5 V vs. Ag/AgCl. LBL assembly decreases the aggregation and increases the loading of PMPDI on the mesoporous ITO substrate, which stabilizes and amplifies the ECL signals. The ECL method exhibits excellent sensitivity and selectivity with good stability and reproducibility, when used to detect dopamine (DA) under optimal experimental conditions.

Sensitive and selective electrochemical determination of uric acid in urine based on ultrasmall iron oxide nanoparticles decorated urchin-like nitrogen-doped carbon.

Hypercrosslinked pyrrole was synthesized via the Friedel-Crafts reaction and then carbonized to obtain urchin-like nitrogen-doped carbon (UNC). Ultrasmall iron oxide nanoparticles were then supported on UNC, and the composite was used to prepare an electrochemical sensor for detecting uric acid (UA) in human urine. FexOy/UNC was characterized and analyzed via scanning electron microscopy, transmission electron microscopy, energy dispersive spectrometry, X-ray diffraction, and X-ray photoelectron spectroscopy. A glassy carbon electrode (GCE) modified with FexOy/UNC was used as an electrochemical sensor to effectively identify UA. The electrochemical behavior of the FexOy/UNC-based UA sensor was studied using differential pulse stripping voltammetry, and the optimal conditions were determined by changing the amount of FexOy/UNC, pH of the buffer solution, deposition potential, and deposition time. Under optimal conditions, the FexOy/UNC-based electrochemical sensor detected UA in the range of 2-200 µM, where the limit of detection (LOD) for UA was 0.29 µM. Anti-interference experiments were performed, and the sensor was applied to the actual analysis of human urine samples. Urea, glucose, ascorbic acid, and many cations and anions present at 100-fold concentrations relative to UA did not strongly interfere with the response of the sensor to UA. The FexOy/UNC electrochemical sensor has high sensitivity and selectivity for uric acid in human urine samples and can be used for actual clinical testing of UA in urine.

Atom-dispersed copper and nano-palladium in the boron-carbon-nitrogen matric cooperate to realize the efficient purification of nitrate wastewater and the electrochemical synthesis of ammonia.

Electrochemical nitrate reduction reaction (NIRR) driven by sustainable energy is not only expected to realize the green production of ammonia under ambient conditions, but also a promising way to purify nitrate wastewater. The ammonia yield rate and Faradaic efficiency of NIRR catalyzed by Pd10Cu/BCN constructed with structural constraints and pre-embedded reducing agent strategies were as high as 102,153 µg h-1 mgcat.-1 and 91.47%, respectively. Pd10Cu/BCN can remove nearly 100% of 50 mg L-1 NO3- without NO2- residue within 10 h, and the realization of this effect does not require the participation of any chloride. Control experiments and DFT calculations explain the efficient operation mechanism of NIRR on Pd10Cu/BCN, where the Pd and CuN4 sites play the role of synergistic catalysis. Compared with the reported literature, Pd10Cu/BCN with good biocompatibility has become an outstanding representative of NIRR catalyst, which provides an alternative way for the green production of ammonia and the purification of nitrate wastewater.