A Chelation-enhanced Fluorescence Assay using Thiourea Capped Carbonaceous Fluorescent Nanoparticles for As (III) Detection in Water Samples.

Herein, we designed a sensitive and selective "Turn-On" fluorescence nanosensor using water-soluble carbonaceous fluorescent nanomaterials (CFNs) functionalized with thiourea (CFNs-Thiourea) for efficient detection of trace concentrations of arsenic (III) in aqueous samples. The CFNs and CFNs-Thiourea were characterized by transmission electron microscopy (TEM), UV-visible spectroscopy (UV-vis) and fourier transformed infrared spectroscopy (FTIR). The emission peak intensity of proposed nanosensor at 425 nm was gradually enhanced on arsenite addition in a wide detection range (3.3-828.5 µg L-1) attributed to the binding of arsenite species with sulfur groups of CFNs-Thiourea. The limit of detection (LOD) was 0.48 µg L-1 being much lower than the World Health Organization (WHO) recommended threshold value of 10 µg L-1. Furthermore, the as-prepared CFNs-Thiourea exhibited a superb selectivity for As (III) compared to various cations and anions, such as; NO3-, NO2-, F-, Ni2+, Fe3+, Cu2+, Ca2+, Mg2+, Zn2+, Fe2+, Hg2+, Pb2+, F-, Cl-, Mn2+, Cr3+, Co2+, Cd2+, Bi3+, Al3+ and As (V) at 100 folds concentration of As (III). The turn on fluorescence nanosensor was successfully exploited for quantification of arsenic in spiked water samples with acceptable efficiencies.

Understanding the bulk and interfacial structures of ternary and binary deep eutectic solvents with a constant potential method: a molecular dynamics study.

In the last decade, deep eutectic solvents (DESs) have emerged as promising electrolytes in supercapacitors and rechargeable batteries due to their unique properties, wide electrochemical windows, low viscosity, and high ionic conductivity. The molecular structural behavior of these solvents, which plays an important role in their efficiency is not deeply understood. Therefore, in this work, by considering two types of DES electrolytes, we investigate their bulk and interfacial structures at the molecular level using molecular dynamics studies. In this regard, two different DESs-a binary DES including choline chloride and urea in a 1 : 2 molar ratio, and a ternary DES containing choline chloride, urea, and ethylene glycol with a molar ratio of 1 : 1 : 2-are considered. For the bulk phase, the partial site-site and center of mass radial distribution functions (RDFs), the mean square displacement (MSD), and the self-diffusion coefficient of the ternary system are explored. We demonstrate that in deep eutectic solvents, in addition to hydrogen-bonding and long-range and short-range correlations, a variety of neutral species play crucial roles in the properties of the bulk phase. Furthermore, considering two graphene sheets as electrodes on both sides of the DES samples, the profiles of the number density, charge density, orientational order parameter, and electrostatic potentials at different potential conditions near the electrodes are investigated. The results reveal the presence of multilayers of the neutral species in the vicinity of electrodes in addition to the ionic components of both DES systems. Finally, the computed differential capacitances (Cd) for DESs disclose that the positive electrode capacitance is higher than that of the negative electrode, and in the ternary system, the total capacitance is greater than in the binary system. Our findings give a better perspective of a new generation of electrolytes at the molecular level for electric double-layer capacitors.

Carbon dots hybrid for dual fluorescent detection of microRNA-21 integrated bioimaging of MCF-7 using a microfluidic platform.

BACKGROUND: MicroRNAs have short sequences of 20 ~ 25-nucleotides which are similar among family members and play crucial regulatory roles in numerous biological processes, such as in cell development, metabolism, proliferation, differentiation, and apoptosis. RESULTS: We reported a strategy for the construction of a dual-emission fluorescent sensor using carbon dots (CDs) and confirmed their applications for ratiometric microRNA-21 sensing and bioimaging of cancer cells in a microfluidic device. The composition of blue CDs (B-CDs) and yellow CDs (Y-CDs) depicts dual-emission behavior which is centered at 409 and 543 nm under an excitation wavelength of 360 nm. With increasing microRNA-21 concentration, the robust and specific binding of DNA probe functionalized B-CDs to complementary microRNA-21 target induced perturbations of probe structure and led to changing fluorescence intensity in both wavelengths. Consequently, the ratio of turn-on signal to turn-off signal is greatly altered. With monitoring of the inherent ratiometric fluorescence variation (ΔF540nm/ΔF410nm), as-prepared BY-CDs were established as an efficient platform for ratiometric fluorescent microRNA-21 sensing, with a wide linear range of 0.15 fM to 2.46 pM and a detection limit of 50 aM. CONCLUSIONS: Furthermore, the proposed assay was applied for detecting microRNA-21 in dilute human serum samples with satisfactory recovery and also in MCF-7 cell lines in the range 3000 to 45,000 (cell mL-1) with a detection limit (3 cells in 10 µL), demonstrating the potential of the assay for clinic diagnosis of microRNA-associated disease. More importantly, the images revealed that MCF-7 cells well labeled with BY-CDs could exhibit the applicability of the proposed microfluidic system as an effective cell trapping device in bioimaging.

Ultrasensitive molecularly imprinted fluorescence sensor for simultaneous determination of CA125 and CA15-3 in human serum and OVCAR-3 and MCF-7 cells lines using Cd and Ni nanoclusters as new emitters.

In the clinical diagnosis of tumors, a single-marker immunoassay may lead to false results. Thus there is a need for an effective and valid method for the simultaneous measurement of multiple tumor markers. In this work, an efficient fluorescence immunosensor for the simultaneous measurement of CA125 and CA15-3 tumor markers was fabricated by utilizing the high selectivity of magnetic molecularly imprinted polymers (MMIPs) and the high sensitivity of a fluorescence (FL) method. Ni nanoclusters (Ni NCs) and noble Cd nanoclusters (Cd NCs) were introduced as efficient and economic emitters, and magnetic graphene oxide (GO-Fe3O4) was applied as a support material for surface molecularly imprinted polymers. Under the most favorable experimental conditions, the fluorescence intensity of the Cd NCs and Ni NCs gradually increased with increasing concentration of CA125 and CA15-3 antigens at a range of 0.0005-40 U mL-1, respectively, with a limit of detection (LOD) of 50 µU mL-1. The developed method had excellent properties including a broad linear range, good reproducibility, and simple operation for the clinical diagnosis of CA 125 and CA 15-3 tumor markers. This molecularly imprinted fluorescence sensor has the potential to be an effective clinical tool for the timely screening of breast cancer in human serum samples and OVCAR-3 and MCF-7 cell lines, and can be applied in clinical diagnostics.

Multienzymes activity of metals and metal oxide nanomaterials: applications from biotechnology to medicine and environmental engineering.

With the rapid advancement and progress of nanotechnology, nanomaterials with enzyme-like catalytic activity have fascinated the remarkable attention of researchers, due to their low cost, high operational stability, adjustable catalytic activity, and ease of recycling and reuse. Nanozymes can catalyze the same reactions as performed by enzymes in nature. In contrast the intrinsic shortcomings of natural enzymes such as high manufacturing cost, low operational stability, production complexity, harsh catalytic conditions and difficulties of recycling, did not limit their wide applications. The broad interest in enzymatic nanomaterial relies on their outstanding properties such as stability, high activity, and rigidity to harsh environments, long-term storage and easy preparation, which make them a convenient substitute instead of the native enzyme. These abilities make the nanozymes suitable for multiple applications in sensing and imaging, tissue engineering, environmental protection, satisfactory tumor diagnostic and therapeutic, because of distinguished properties compared with other artificial enzymes such as high biocompatibility, low toxicity, size dependent catalytic activities, large surface area for further bioconjugation or modification and also smart response to external stimuli. This review summarizes and highlights latest progress in applications of metal and metal oxide nanomaterials with enzyme/multienzyme mimicking activities. We cover the applications of sensing, cancer therapy, water treatment and anti-bacterial efficacy. We also put forward the current challenges and prospects in this research area, hoping to extension of this emerging field. In addition to therapeutic potential of nanozymes for disease prevention, their practical effects in diagnostics, to monitor the presence of SARS-CoV-2 and related biomarkers for future pandemics will be predicted.

CuO/Cu-MOF nanocomposite for highly sensitive detection of nitric oxide released from living cells using an electrochemical microfluidic device.

The integration of large surface area and high catalytic profiles of Cu-MOF and CuO nanoparticles is described toward electrochemical sensing of nitric oxide (NO) in a microfluidic platform. The CuO/Cu-MOF nanocomposite was prepared through hydrothermal method, and its formation was confirmed by scanning electron microscopy (SEM), transmission electron microscopy (TEM), X-ray powder diffraction (XRD), X-ray photoelectron spectroscopy (XPS), and energy-dispersive X-ray spectroscopy (EDS). The CuO/Cu-MOF nanostructured modified Au electrodes enabled electrocatalytic NO oxidation at 0.6 V vs. reference electrode, demonstrating linear response over a broad concentration range of 0.03-1 µM and 1-500 µM with a detection limit of 7.8 nM. The interference effect of organic molecules and common ions was negligible, and the sensing system demonstrated excellent stability. Finally, an electrochemical microfluidic NO sensor was developed to detect of NO released from cancer cells, which were stimulated by L-arginine. Furthermore, in the presence of Fe3+, the stressed cells produced more NO. This work offers considerable potential for its practical applications in clinical diagnostics through determination of chemical symptoms in microliter-volume biological samples. Electrochemical microfluidic NO sensor was developed for detection of NO released from cancer cells. This miniaturized device consumes less materials and provides the basis for greener analytical chemistry.

A strategy for visual optical determination of glucose based on a smartphone device using fluorescent boron-doped carbon nanoparticles as a light-up probe.

Boronic acid-doped carbon nanoparticles were prepared and are shown to undergo aggregation induced emission (AIE). The nanoparticle composite is a viable fluorescent probe for glucose determination by using the RGB technique and a smartphone. The structure and the chemical composition of the doped carbon nanoparticles were confirmed by SEM, TEM, FTIR and UV-vis spectroscopy. The combination of 4-carboxyphenylboronic acid with o-phenylenediamine and rhodamine B endowed the hybrid with high fluorescence intensity (quantum yield 46%). Compared with conventional two-step preparation of boronic acid-based fluorescent probes for glucose, the present one step synthesis strategy is simpler and more effective. The addition of glucose causes the formation of covalent bonds between the cis-diols group of glucose molecules and boronic acid moiety. Fluorescent intensity can be quantified using dual wavelengths simultaneously, where both increases, as the target analytes bind to the bronic acid. These variations was monitored by the smartphone camera, and the green channel intensities of the colored images were processed by using the RGB option of a smartphone. The assay works in the 32 µM to 2 mM glucose concentration range and has an 8 µM detection limit. The method was successfully used for the assay of glucose in diluted human serum. Graphical abstractThe fluorometric method was developed for determination of glucose using boron doped carbon nanoparticles (BCNBs). The BCNPs aggregate after covalent binding between the cis-diols of glucose and boronic acid. The green channel of the images is recorded by a smartphone camera.

Functionalized fluorescent carbon nanostructures for targeted imaging of cancer cells: a review.

This short review (with 72 refs.) summarizes the state of the art in fluorometric methods for targeted imaging of cancer cells and tumor tissues in order to differentiate between normal cells and cancer cells. Following an introduction into the field and after presenting an overview on the most commonly used carbon dots and graphene quantum dots, we describe methods based on peptide based targeting, aptamer based targeting, antibody based targeting, and ligand-based targeting. A concluding section summarizes the current state and challenges, and discusses future perspectives. Graphical abstract An overview is given on the applications of carbon dots (CDs) in target-specific imaging and differentiation of cancerous cells from normal cells. Several classes of ligands (including aptamers, peptides, antibodies), especially small molecules (such as FA)) have been reported for functionalizing of CDs.

Ratiometric enhanced fluorometric determination and imaging of intracellular microRNA-155 by using carbon dots, gold nanoparticles and rhodamine B for signal amplification.

An ultrasensitive and highly reliable ratiometric assay is described for the determination of microRNA-155. It works at the attomolar concentration level and has high selectivity which warrants its potential application in cancer biomarker tracking. The excellent performance of this method results from (a) the use of a hybrid conjugate prepared from Rhodamine B (RhB), carbon dots (CDs) and probe-microRNA, and (b) from the measurement of fluorescence resonance energy transfer (FRET) that is observed in the AuNP/target-microRNA system as a result of RNA hybridization. The dye RhB (emission peak at 580 nm) serves as an internal reference. The sensitivity of this assay is increased by about 30% because of the broad emissions of CDs (489 nm and 665 nm) through a sequential FRET phenomenon. RhB-CDs were covalently bio-conjugated to probe microRNA. In the presence of AuNPs, the fluorescence of the CDs is quenched, while in the presence of microRNA-155, the ratio of fluorescences at 489 and 665 nm (I489/I665) is enhanced again. A linear relationship exists between the ratio of fluorescence and the concentration of microRNA-155 in the range from 1 aM to 0.1 µM, and the detection limit is 0.3 aM. The assay was applied to quantitative studies of target microRNA-155 in multiple pathways associated with cancer progression in biological fluids include human serum samples and cancer cells. The nanoprobe also deliver clear signal to microRNA target in fixed and lived MDA-MB-231 cells. Graphical abstract A ratiometric FRET sensing method used for microRNA-155 detection at aM concentration level using CDs and AuNPs as donor-acceptor respectively and Rhodamine B as amplification reagent. The application of assay for imaging of microRNA-155 in fixed and live MDA-MB-231 cells is demonstrated.

A FRET immunosensor for sensitive detection of CA 15-3 tumor marker in human serum sample and breast cancer cells using antibody functionalized luminescent carbon-dots and AuNPs-dendrimer aptamer as donor-acceptor pair.

We proposed a FRET immunosensing for detection of CA15-3 tumor marker by highly biospecific interactions between CA 15-3 antigen and the corresponding antibody and aptamer. In this sandwich type immunoassay, CA15-3 antibody-functionalized carbon dots and AuNPs labeled PAMAM-Dendrimer/aptamer were used as donor/acceptor, respectively. When CA 15-3 Ag was added to homogenous immunoassay, the strong complex interaction between CA15-3 Ab-CA15-3 Ag- aptamer caused in more coming closer carbon dot and AuNPs and more decreasing fluorescence signal. The decreased fluorescence intensity was linear at three ranges including in concentration range 1.1 µUmL-1 to 16 µU mL-1 with regression of R2 = 0.9879, at the concentration range 16 µU mL-1 to 0.163 mU mL-1 with regression of R2 = 0.9944 and at the concentration range 0.163 mU mL-1 to 5.0 mU mL-1 with regression of R2 = 0.9805. The detection limit of the FRET immunoassay was 0.9 µU mL-1. This assay revealed good sensitivity and specificity with MDA-MB-231 breast cancer cells concentrations from 1000 to 40000 cells/mL with correlation coefficient of 0.9955 and detection limit of 300 cells/mL (3 cells in 10 µL of injected sample). In addition, this FRET immunosensing is applicable in diluted human serum. The recovery values were in the range of 95.86-96.97% for CA 15-3 Ag in spiked serum sample with RSD lower than 7.3%. The proposed immunoassay could be a valid model for establishing other immunoassays for detection of different cancer tumor markers with relevant antigens and antibodies.

Ni-hemin metal-organic framework with highly efficient peroxidase catalytic activity: toward colorimetric cancer cell detection and targeted therapeutics.

BACKGROUND: Given the great benefits of artificial enzymes, a simple approach is proposed via assembling of Ni2+ with hemin for synthesis of Ni-hemin metal-organic-frameworks (Ni-hemin MOFs) mimic enzyme. The formation of the Ni-hemin MOFs was verified by scanning electron microscopy, Transmission electron microscopy, X-ray powder diffraction, X-ray photoelectron spectroscopy, Fourier transform infrared spectroscopy, Energy-dispersive X-ray spectroscopy and UV-vis absorption spectroscopy. This novel nanocomposite exhibited surprising peroxidase like activity monitored by catalytic oxidation of a typical peroxidase substrate, 3,3,5,5'-tetramethylbenzidine, in the presence of H2O2. By using folic acid conjugated MOF nanocomposite as a recognition element, we develop a colorimetric assay for the direct detection of cancer cells. RESULTS: The proposed sensor presented high sensitivity and selectivity for the detection of human breast cancer cells (MCF-7) and Human Caucasian gastric adenocarcinoma. By measuring UV-vis absorbance response, a wide detection range from 50 to 105 cells/mL with a detection limit as low as 10 cells/mLwas reached for MCF-7 cells. We further discuss therapeutics efficiency of Ni-hemin MOFs in the presence of H2O2 and ascorbic acid. Peroxidase-mimic Ni-hemin MOFs as reactive oxygen species which could damage MCF-7 cancer cells, however for normal cells (human embryonic kidney HEK 293 cells) killing effect was negligible. CONCLUSIONS: Based on these behaviors, the developed method offers a fast, easy and cheap assay for the interest in future diagnostic and treatment application.

Fluorometric determination of microRNA-155 in cancer cells based on carbon dots and MnO2 nanosheets as a donor-acceptor pair.

A fluorometric method is presented for sensitive deternination of microRNA. It is making use of carbon dots (C-dots) loaded with a DNA probe as fluorophore and MnO2 nanosheets as the quenching agent. The blue-green fluorescence of the DNA-loaded C-dots is quenched by the MnO2 nanosheets, but restored on binding target microRNA-155. The maximum excitation wavelength and the maximum emission wavelength of C-dots are at 360 nm and 455 nm, respectively. Fluorescence correlates linearly with the log of the microRNA-155 concentration in two ranges, viz. from 0.15 to 1.65 aM and from 1.65 to 20 aM. The detection limit is as low as 0.1 aM. The assay can discriminate between fully complementary and single-base mismatch microRNA. The assay displayed high specificity when used to detect MCF-7 breast cancer cells which can be detected in concentrations from 1000 to 45,000 cells·mL-1, with a 600 cells·mL-1 detection limit. The method was applied to the analysis of serum samples spiked with microRNA, and satisfactory results were acquired. Graphical abstract Schematic of a fluorometric sensing platform for miRNA-155. The method relies on a FRET process between C-dots and MnO2 nanosheets. This strategy has a practical application for detection of miRNA in cell lines and biological fluids.

Electrochemical pretreatment of amino-carbon nanotubes on graphene support as a novel platform for bilirubin oxidase with improved bioelectrocatalytic activity towards oxygen reduction.

The electrochemical conditioning of amino-carbon nanotubes (CNTs) on a graphene support in an alkaline solution is used to produce -NHOH as hydrophilic functional groups for the efficient immobilization of bilirubin oxidase enzyme. The application of the immobilized enzyme for the direct electrocatalytic reduction of O2 is investigated. The onset potential of 0.81 V versus NHE and peak current density of 2.3 mA cm(-2) for rotating modified electrode at 1250 rpm, indicate improved biocatalytic activity of the proposed system for O2 reduction.

Highly sensitive electrochemical aptasensor for immunoglobulin E detection based on sandwich assay using enzyme-linked aptamer.

Here, we describe the fabrication of an electrochemical immunoglobulin E (IgE) aptasensor using enzyme-linked aptamer in the sandwich assay method and thionine as redox probe. In this protocol, 5'-amine-terminated IgE aptamer and thionine were covalently attached on glassy carbon electrode modified with carbon nanotubes/ionic liquid/chitosan nanocomposite. Furthermore, another IgE aptamer was modified with biotin and enzyme horseradish peroxidase (HRP), which attached to the aptamer via biotin-streptavidin interaction. Electrochemical impedance spectroscopy (EIS) and cyclic voltammetry were performed at each stage of the chemical modification process to confirm the resulting surface changes. The presence of IgE induces the formation of a double aptamer sandwich structure on the electrode, and the electrocatalytic reduction current of thionine in the presence of hydrogen peroxide was measured as the sensor response. Under optimized conditions and using differential pulse voltammetry as the measuring technique, the proposed aptasensor showed a low detection limit (6 pM) and high sensitivity (1.88 µA nM(-1)). This aptasensor also exhibited good stability and high selectivity for IgE detection without an interfering effect of some other proteins such as bovine serum albumin (BSA) and lysozyme. The application of the aptasensor for IgE detection in human serum sample was also investigated. The proposed protocol is quite promising as an alternative sandwich approach for various protein assays.

Magnetic N-doped carbon derived from mixed ligands MOF as effective electrochemiluminescence coreactor for performance enhancement of SARS-CoV-2 immunosensor.

COVID-19 as an infectious disease with rapid transmission speed is caused by severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2), so, early and accurate diagnostics of COVID-19 is quite challenging. In this work, the selective and sensitive self-enhanced ECL method to detect of SARS-CoV-2 protein was designed with magnetic N-doped carbon derived from dual-ligand metal-organic frameworks (MOF) (CoO@N-C) with the primary and tertiary amino groups as a novel coreactant that covalently combined with Ru(bpy)2(phen-NH2)2+ as electrochemiluminescence (ECL) emitter. Mixed-ligand strategy and selected nitrogen-containing ligands, 4,4',4''-((1,3,5-triazine-2,4,6-triyl) tris-(azanediyl)) tribenzoic acid (H3TATAB) with 2-aminoterephthalic acid (BDC-NH2) were used for synthesis of the proposed MOF. Also, magnetic CoO@N-C with high synergistically charge transfer kinetics and good stability can be used as an effective platform/coreactor on the ITO electrode which load more Ru-complex as signal producing compound and SARS-CoV-2 N protein antibody to increase the sensitivity of the immunosensor. Furthermore, (CoO@N-C) as coreactor improved the ECL signal of the Ru (II)-complex more than 2.1 folds compared to tripropylamine. In view of these competences, the novel "on-off" ECL biosensor performed with great stability and repeatability for detection of SARS-CoV-2 protein, which exhibited a broad linearity from 8 fg. mL-1 to 4 ng. mL-1 (6 order of magnitude) and an ultra-low limit of detection 1.6 fg. mL-1. Finally, this proposed method was successfully applied to detect of SARS-CoV-2 N protein in serum sample with satisfactory results, indicating the proposed immunosensor has the potential for quick analysis of SARS-CoV-2.

Electrochemical monitoring of hydrogen peroxide by a signal-amplified microfluidic chip coupled with colloidal VO2 nanostructures as a peroxidase enzyme mimic.

We present a novel electrochemical microfluidic device for the sensitive and selective detection of hydrogen peroxide (H2O2) through a VO2 nanostructure enzyme mimic. The low-cost ($0.50) microfluidic chip was fabricated using a simple and rapid prototyping technique via three syringe needles. Each needle played the role of an electrode (working, reference, and counter), and was connected by micro-hoses to a construction of the electrochemical microfluidic chip. The colloidal VO2 nanoflakes with peroxidase-like activity could be easily transferred on to the electrodes by a syringe, for development of a novel electrochemical platform to enable the detection of H2O2. The unique microfluidic electrochemical sensor delivered a wide linear dynamic range from 0.5 to 300 µM, with a limit of detection of 0.14 µM. The facile, rapid, sensitive, and selective as-fabricated H2O2 sensors were proven to be appropriate for the real-time monitoring of H2O2 released from PC12 cells. The integration of a microfluidic sensor with an enzyme mimic nanostructure is essentially a promising strategy for the low-cost and accurate monitoring of physiological processes.

Electrochemical-based remote biomarker monitoring: Toward Internet of Wearable Things in telemedicine.

Internet of Wearable Things (IoWT) will be a major breakthrough for remote medical monitoring. In this scenario, wearable biomarker sensors have been developing not only to diagnose point-of-care (POC) of diseases, but also to continuously manage them. On-body tracking of biomarkers in biofluids is regarded as a proper substitution of conventional biomarker sensors for dynamic sampling and analyzing due to their high sensitivity, conformability, and affordability, creating ever-rising the market demand for them. In a wireless body area network (WBAN), data is captured from all sensors on the body to a smartphone/laptop, and sent the sensed data to a cloud for storing, processing, and retrieving, and ultimately displayed the data on custom applications (Apps). Wearable IoT biomarker sensors are used for early diseases diagnosis and continuous monitoring in developing countries in which people hardly access to healthcare systems. In this review, we aim to highlight a wide range of wearable electrochemical biomarker sensors, accompanied by microfluidics for continuous sampling, which will pave the way toward developing wearable IoT biomarker sensors to track health status. The current challenges and future perspective in skin-conformal biomarker sensors will be discussing their potential applicability for IoWT in cloud-based telemedicine.

Ultrasensitive Ratiometric Fluorescence Bioassay for Accurate Detection of Covid-19-Specific Nucleocapsid Protein in Clinical Serum Samples Using Modified Cleavable Mesoporous SiO2 Satellite-Enriched Carbon Dots.

Due to the presence of various autofluorescent compounds in biological samples like serum and the photobleaching of organic fluorophores, fluorescence sensing has limited practical applicability. This study describes the development of an improved ratiometric fluorescence assay to determine the nucleocapsid protein (N protein), one of the most conserved biomarkers of Covid-19 in spiked and serum samples using highly stable buffer-based near IR-dual emission carbon dots (CDs) encapsulated into the cavities of cleavable silica nanocapsule (SNCs) nanocomposite. The cavities of cleavable silica nanocapsules (SNCs) and the formed core-shell CDs@ SNCs were used as a superior reservoir of fluorescent markers produced by cohydrolyzing tetraethyl orthosilicate and diiminosilane linker, which held hundreds of CDs in silica shell frameworks. The SiO2 nanocomposite was modified with an N protein antibody that specifically paired to the receptor binding region of the Cov-19 spike protein subunit. CDs were taken out of SNCs by NaBH4 reduction, and the released CDs exhibited dual emission at 475 and 675 nm when excited at 400 nm. Ratiometric detection is completed over a binding-induced, concentration-dependent immuno-affinity of the N protein that drives the fluorescence quenching phenomenon between the CDs as fluorophore and the AuNPs as quencher. As the N protein concentration increased, the intensity of the red emission (675 nm) dropped, whereas the intensity of the green emission (475 nm) already remained constant, which is due to sandwich immunoassays of CDs around AuNPs. Using the exceptional fluorescent characteristics of CDs and the high selectivity of nanocomposite functionalized with N-protein antibody, the developed assay efficiently eliminates the autofluorescence background interference of serum samples. The fluorescence ratio (I475/I675) provides a limit of detection of 2 pg mL-1 over a linear range of 0.01 to 5 ng mL-1 and exhibits an amplified sensitivity of 54 times compared to conventional immunoassay using CDs as fluorescent labels. With one-step signal amplification and requiring small sample quantities (only 20 µL), this sensing platform can be effectively used for the accurate detection of N protein, and no cross-reactivity is detected in the presence of different interfering agents.

A novel highly sensitive compilation-detachment fluorescence sensing strategy based on RNA-cleavage DNAzyme for MDA-MB-231 breast cancer biomarker determination.

Herein, we designed a novel and highly sensitive fluorescence multicomponent detachable platform for MDA-MB-231 breast cancer cell detection as a model. The RNA cleavage DNAzyme was used as a central operator of the multicomponent probe through which compilation and induced detachment of probe was done. During the compilation step, the dsDNA-Sybr green 1 complexes on gold nanoparticles (GNP@dsDNA@SG1) were assembled. The intercalated Sybr green in the DNA structure has been used as an amplified signal generator on one site of DNAzyme and magnetic nanoparticles (MNP) act as a biological carrier and probe collector on the opposite side. The enzyme activator co-factor (MDA-MB-231 cell cytoplasmic protein) provokes the activation of the catalytic core of enzyme sequence in the DNAzyme molecule, followed by cleavage reaction in the substrate sequence and releasing GNP@ dsDNA@SG1 into the solution. The results indicate that the Sybr green emission fluorescence (520 nm) increases with the increment of MDA-MB-231 protein concentration in the linear dynamic range of 8.10 × 10-2 to 1.95 ng ml-1 (0.77 × 10-3-0.019 cell ml-1) with a detection limit (LOD) of 1/72 × 10-2 pg ml-1 under optimal conditions. The proposed immunosensor has great potential in developing ultrasensitive and rapid diagnostic platforms.

Ultrasensitive immunosensor for monitoring of CA 19-9 pancreatic cancer marker using electrolyte-gated TiS3 nanoribbons field-effect transistor.

Measuring CA 19-9 antigen level is critical for early diagnosis of pancreatic cancer, monitoring the treatment process, and predicting disease recurrence. The purpose of this research is to assess the application of novel few-layered TiS3 nanoribbons material as a channel material in electrolyte-gated field-effect transistor immunosensor for rapid detection of CA 19-9 antigen as a cancer marker. Accordingly, TiS3 nanoribbons were produced through liquid-phase exfoliation of as-synthesized TiS3 whiskers in N, N-dimethylformamide. Then, dispersed TiS3 nanoribbons were drop cast onto the FET surface to form an active channel material between source and drain electrodes. Subsequently, the channel surface was modified by utilizing 1-naphthylamine (NA) and glutaraldehyde (GA) to strengthen the binding of monoclonal antibody 19-9 to TiS3 nanoribbons. Spectroscopic and microscopic methods were utilized for comprehensive characterizations. Electrical characterization of electrolyte-gated TiS3 nanoribbons field-effect transistor represented a depletion-mode n-type behavior with field-effect mobility of 0.059 cm2/Vs, current on/off ratio of 10.88 and subthreshold swing (SS) of 450.9 mV/decade. With increasing in CA 19-9 antigen concentration from 1.0 × 10-12 U/mL to 1.0 × 10-5 U/mL, a decrease in the drain current occurred with high sensitivity of 0.04 µA/decade and a detection limit of 1.3 × 10-13 U/mL. Additionally, the proposed TiS3 nanoribbons FET immunosensor exhibited outstanding selectivity, and its good performance was compared with an enzyme-linked immunosorbent assay (ELISA) for spiked real human serum samples. The good and satisfactory obtained results of the proposed immunosensor suggest that the developed platform can be a superb candidate for cancer diagnosis and therapeutic monitoring.