Ti3C2 MXene-based nanozyme as coreaction accelerator for enhancing electrochemiluminescence of glucose biosensing.

Delamination of the exfoliated multilayer MXenes with electro-catalysts, not only leads to increasing surface area for high electrochemiluminescent (ECL) signal tracer loading but also provides highly sensitive achievements in a coreaction accelerator manner. To this end, herein, we used bromophenol blue (BPB)-delaminated multilayer Ti3C2 MXene as both a coreaction accelerator to promote the electrochemiluminescent (ECL) reaction rate of luminol (LUM) and the co-reactant H2O2 and a substrate for retaining high loading of glucose oxidase (GOx)-conjugated polyethylene imine (PEI) along with luminophore species into more open structure of Ti3C2 MXene for sensitive detection of glucose. In the presence of glucose, in situ generating H2O2 product through a GOx-catalyzed process could produce abundant •OH radicals via the peroxidase-like activity of the BPB@Ti3C2 in the LUM ECL reaction. Moreover, decreasing the distance between the high-content LUM into the BPB@Ti3C2 and the generated •OH, minimizes the decomposition of highly active •OH, providing a superb ECL signal. Last, the proximity of incorporated GOx into the delaminated Ti3C2 MXene near the electrode allows efficient electron transfer between the electrode and enzyme. The integration of such amplifying effects endowed high sensitivity and excellent selectivity for glucose with a low limit of detection of 0.02 µM in the wide range of 0.01 µM-40,000 µM, enabling the feasibility of the glucose analysis in human serum samples. Overall, the enhanced ECL based on the BPB@Ti3C2 opens a new horizon to develop highly sensitive MXene-based ECL toward the field of biosensors.

A label-free multicolor colorimetric and fluorescence dual mode biosensing of HIV-1 DNA based on the bifunctional NiFe2O4@UiO-66 nanozyme.

Finding the DNA of the human immune deficiency virus (HIV) with simple and sensitive detection is the main challenge in early diagnosis of AIDS. Herein, two-point separation strategies based on the colorimetric and fluorescence are introduced. The naked-eye qualitative and semiquantitative colorimetric, and also accuracy fluorescence quantification of HIV-1 DNA were applied using label-free NiFe2O4@UiO-66 nanozyme with both functions of peroxidase-mimetic like and emitting fluorescence. The DNA probe-conjugated nanozyme is employed to hybridize a sequence of HIV-1. NiFe2O4@UiO-66 nanozymes catalyze the decomposition of H2O2 to •OH which can produce a remarkable fluorescent product 2-hydroxyterephthalic acid (TAOH) by the oxidation of the bridging ligand of weakly fluorescent terephthalic acid (TA). The accessibility of H2O2 toward confined-NiFe2O4 MNPs was reduced by increasing the HIV-1 target DNA concentration, resulting in the fluorescence intensity of TAOH being decreased. Meanwhile, remaining the unreacted H2O2 was transferred an acidic colorimetric solution containing FeSO4 and gold nanorods (AuNRs). Increasing the amount of H2O2 available for longitudinal etching of AuNRs due to •OH-generating Fe+2-catalyzed H2O2 is reponsible for different colors from brownish to colorless depending on the HIV-1 target DNA concentration. The fluorescence intensity and obtained colors have offered the sensitive biosensing methods with a linear range from 0.05 to 300 and 1-200 pM, respectively with a detection limit as low as 1 fM. Our study revealed that the applied sensing assay provides a cost-effective and straightforward qualitative, semiquantitative, and sensitive quantitation visible monitoring without the necessity of high-end instruments for HIV-1 detection in a human blood plasma/serum samples.

Smartphone-Based Electrochemiluminescence for Visual Simultaneous Detection of RASSF1A and SLC5A8 Tumor Suppressor Gene Methylation in Thyroid Cancer Patient Plasma.

Visual one-step simultaneous detection of low-abundance methylation is a crucial challenge in early cancer diagnosis in a simple manner. Through the design of a closed split bipolar electrochemistry system (BE), detection of promoter methylation of tumor suppressor genes in papillary thyroid cancer, RASSF1A and SLC5A8, was achieved using electrochemiluminescence. For this purpose, electrochemiluminescence of luminol loaded into the Fe3O4@UiO-66 and gold nanorod-functionalized graphite-like carbon nitride nanosheet (AuNRs@C3N4 NS), separately, on the anodic and cathodic pole bipolar electrodes (BPEs) in two different chambers of a bipolar cell were recorded on a smartphone camera. To provide the same electric potential (ΔEelec) through the BPEs to conduct simultaneous light emission, as well as to achieve higher sensitivity, anodic and cathodic poles BPEs were separately connected to ruthenium nanoparticles electrodeposited on nitrogen-doped graphene-coated Cu foam (fCu/N-GN/RuNPs) to provide a hydrogen evolution reaction (HER) and polycatechol-modified reduced graphene oxide/pencil graphite electrode (PC-rGO/PGE) to provide electrooxidation of hydrazine. Moreover, taking advantages of the strong cathodic ECL activity due to the roles of AuNRs, as well as the high density of capture probes on the UiO-66 and Fe3O4 roles in improving the signal-to-background ratio (S/B) in complicated plasma media, a sensitive visual ECL immunosensor was developed to detect two different genes as model target analytes in patient plasma samples. The ability of discrimination of methylation levels as low as 0.01% and above 90% clinical sensitivity in thyroid cancer patient plasma implies that the present strategy is able to diagnose cancer early, as well as monitor responses of patients to therapeutic agents.

Electrochemical immunosensor for determination of cardiac troponin I using two-dimensional metal-organic framework/Fe3O4-COOH nanosheet composites loaded with thionine and pCTAB/DES modified electrode.

Cardiac troponin-I (CTnI) is one of the most popular biomarkers which can be utilized for the diagnosis and control of acute myocardial infarction in clinical practice. Here, a sandwich-type electrochemical immunosensor has been established using the zinc-based metal-organic framework/Fe3O4-COOH/thionine labeled anti-CTnI monoclonal antibody (Ab1-Zn-MOF/Fe3O4-COOH/Thi) nanocomposite as signaling molecule and a polymer film of cetyltrimethylammonium bromide (pCTAB) in the presence of choline chloride-urea deep eutectic solvent (DES) and anti-CTnI polyclonal antibody (Ab2) as immobilization substance of detecting surface. The porous ultrathin layers of Zn-MOF nanosheets successfully prepare a well-defined structure for Fe3O4-COOH electrocatalyst and Thi within a certain two dimensional (2D) regions, which enhances electrochemical reduction of Thi. The Ab1-Zn-MOF/Fe3O4-COOH/Thi nanocomposites were introduced to CTnI in the specimen and on the surface of pCTAB/DES-Au-SPE quantitative determination of CTnI was achieved using differential pulse voltammetry after sandwiching the CTnI target between Ab1-nanocomposite and Ab2 which was encapsulated into the pCTAB/DES-Au-SPE. This immunosensor indicated the appropriate assay performance for CTnI with the detection range of 0.04 ng mL-1 to 50 ng mL-1 and the limit of detection of 0.0009 ng mL-1. This study provides convenient plan for sensitive detection of bioanalytes and opens a path for the establishment of user-friendly and cost-effective device.

Electrochemiluminescence paper-based screen-printed electrode for HbA1c detection using two-dimensional zirconium metal-organic framework/Fe3O4 nanosheet composites decorated with Au nanoclusters.

Glycated hemoglobin (HbA1c) is one of the most popular biomarkers which can be utilized for the diagnosis and control of diabetes in clinical practice. In this study, a sandwich paper-based electrochemiluminescence (ECL) biosensor has been developed using the zirconium metal-organic framework/Fe3O4(trimethyl chitosan)/gold nanocluster (Zr-MOF/Fe3O4(TMC)/AuNCs) nanocomposite as tracing tag to label anti-HbA1c monoclonal antibody and reduced graphene oxide (rGO) as immobilization platform of sensing element. The screen-printed electrodes (SPEs) were constructed and modified by sputtering a thick layer of gold on the paper substrate, followed by electrochemical reduction of aminophenylboronic acid (APBA)-functionalized GO to rGO/APBA, respectively. Different types of surface analysis methods were applied to characterize the Zr-MOF/Fe3O4(TMC)/AuNCs nanomaterials fabricated. Finally, antibody-labeled Zr-MOF/Fe3O4(TMC)/AuNCs nanocomposites were subjected to HbA1c in the sample and on the paper-based SPE. Quantitative measurement of HbA1c was performed using ECL and cyclic voltammetry (CV) over a potential range of - 0.2 to 1.7 V vs gold reference electrode with a sweep rate of 0.2 V.s-1 in the presence of triethylamine as a co-reactant after sandwiching the HbA1c target between antibody and APBA on the sensing area. This immunosensor demonstrated the desirable assay performance for HbA1c with a wide response range from 2 to 18% and a low detection limit (0.072%). This new strategy provides an effective method for high-performance bioanalysis and opens avenues for the development of high-sensitive and user-friendly device. Graphical abstract.

Electrochemiluminescent biosensor for ultrasensitive detection of lymphoma at the early stage using CD20 markers as B cell-specific antigens.

Herein, by taking advantage of the special binding of an aptamer to the membrane surface of a B cell and accumulation of the positive charges of a nanocomposite, including luminol-chitosan-platinum nanoparticles (L-Cs-Pt NPs), on the negatively charge of the aptamer phosphate backbone, a sensitive, simple, selective and rapid strategy for the detection of lymphoma cells by a new label-free electrogenerated chemiluminescence (ECL) aptasensor has been introduced. With increasing concentrations of B lymphoma cells, the nanocomposite detaches from the aptamer, leading to a decrease in the ECL of a luminol and H2O2 system. With high loading of luminol and Pt NPs on a chitosan, together with the electrocatalytic effect of Pt NPs, enhanced sensitive detection of cancer cells with a limit of detection of 31 cells/mL was achieved. Step-by-step modification and biosensor response to cancer cells was monitored by electrochemical impedance spectroscopy (EIS), cyclic voltammetry (CV), and ECL. The aptasensor exhibited excellent specificity for lymphoma cells vs breast cancer (MCF-7) and human embryonic kidney (HEK293) cell lines as potential interferents. Finally, the performance of the aptasensor in blood samples was assessed against a commercial flow cytometric method. Satisfactory results confirmed the applicability of the proposed biosensing platform.

Point-of-care biosensors in medicine: a brief overview of our achievements in this field based on the conducted research in EMRI (endocrinology and metabolism research Institute of Tehran University of medical sciences) over the past fourteen years.

The growing demand of diagnostic tools with enhanced analytical characteristics in term of sensitivity, selectivity, and low response time has encouraged researches to conduct their research towards development of point-of-care (POC) biosensors. POC diagnostic devices are powerful tools for detection, diagnosis, and monitoring of diseases at its initial stage. The above characteristics encouraged us to conduct active multidisciplinary and collaborative research oriented towards the design and development of POC sensing systems. Here, we present a brief overview of our recent achievement in the field of biomedical POC devices implemented in paper based microfluidic and screen printing electrodes and discuss the critical limitations that need to be surmounted to facilitate their translation into clinical practice in the future.

Electrochemical sensors and biosensors based on the use of polyaniline and its nanocomposites: a review on recent advances.

Polyaniline and its composites with nanoparticles have been widely used in electrochemical sensor and biosensors due to their attractive properties and the option of tuning them by proper choice of materials. The review (with 191 references) describes the progress made in the recent years in polyaniline-based biosensors and their applications in clinical sensing, food quality control, and environmental monitoring. A first section summarizes the features of using polyaniline in biosensing systems. A subsequent section covers sensors for clinical applications (with subsections on the detection of cancer cells and bacteria, and sensing of glucose, uric acid, and cholesterol). Further sections discuss sensors for use in the food industry (such as for sulfite, phenolic compounds, acrylamide), and in environmental monitoring (mainly pesticides and heavy metal ions). A concluding section summarizes the current state, highlights some of the challenges currently compromising performance in biosensors and nanobiosensors, and discusses potential future directions. Graphical abstract Schematic presentation of electrochemical sensor and biosensors applications based on polyaniline/nanoparticles in various fields of human life including medicine, food industry, and environmental monitoring. The simultaneous use of suitable properties polyaniline and nanoparticles can provide the fabrication of sensing systems with high sensitivity, short response time, high signal/noise ratio, low detection limit, and wide linear range by improving conductivity and the large surface area for biomolecules immobilization.

Enhanced Visual Wireless Electrochemiluminescence Immunosensing of Prostate-Specific Antigen Based on the Luminol Loaded into MIL-53(Fe)-NH2 Accelerator and Hydrogen Evolution Reaction Mediation.

A sensitive prostate-specific antigen (PSA) detection method using a visual-readout closed bipolar electrode (BPE) system has been introduced by integration of hydrogen evolution reaction (HER) in cathodic pole and electrochemiluminescence (ECL) of luminol loaded within the MIL-53(Fe)-NH2 (L@MIL-53(Fe)-NH2) in the anodic pole. The cathode of the BPE was electrochemically synthesized by 3D porous copper foam, followed by decorating with nitrogen-doped graphene nanosheet and ruthenium nanoparticles. As an alternative, we employed carboxylate-modified magnetic nanoparticles (MNPs) for immobilization of the primary antibody (Ab1) and utilized the L@MIL-53(Fe)-NH2 conjugated to secondary antibody (Ab2) as a signaling probe and coreaction accelerator. After sandwiching the target PSA between Ab1 and Ab2, the MNP/Ab1-PSA-Ab2/L@MIL-53(Fe) were introduced to a gold anodic BPE. Finally, the resulting ECL of luminol and H2O2 at the anodic poles was monitored using a photomultiplier tube (PMT) or digital camera. The PMT and visual (camera)-based detections showed linear responses from 1 pg mL-1 to 300 ng mL-1 (limit of detection 0.2 pg mL-1) and 5 pg mL-1 to 200 ng mL-1 (limit of detection 0.1 pg mL-1), respectively. This strategy provides an effective method for high-performance bioanalysis and opens a new door toward the development of the highly sensitive and user-friendly device.

Voltammetric immunosensor for E-cadherin promoter DNA methylation using a Fe3O4-citric acid nanocomposite and a screen-printed carbon electrode modified with poly(vinyl alcohol) and reduced graphene oxide.

Silencing of tumor suppressor genes (E-cadherin) by promoter DNA methylation may lead to the development of invasive phenotypes in epithelial tissues. The authors describe an electrochemical nanobiosensor for early detection and screening of circulating methylated DNA as a biomarker for cancers. First, the antibody against 5-methylcytosine was physically immobilized onto modified with reduced graphene oxide and polyvinylalcohol. In the next step, methylated target DNA in samples was hybridized with ssDNA probe conjugated to Fe3O4-citric acid nanocomposites and placed on the modified electrode. Then, the hexacyanoferrate redox system was added and electron transfer recorded. Cyclic voltammetry and electrochemical impedance spectroscopy showed that the modification process was well accomplished. Quantitative measurement of E-cadherin DNA promoter methylation was performed using differential pulse voltammetry. The electrochemical analysis achieved in the presence and absence of nonmethylated DNA mixed with samples indicated the high specificity and selectivity in methylation analysis using this system. With the linear range of concentration from 1 × 10-4 ng.mL-1 to 20 ng.mL-1 and the detection limit of 9 × 10-5 ng.mL-1, this method represents a promising approach for analysis of other biomarkers. Graphical abstract A label free electrochemical nanobiosensor was constructed for detection of methylated circulating cell-free DNA using screen-printed carbon electrode (SPCE) modified with reduced graphene oxide (rGO) and polyvinylalcohol (PVA).

Visual electrochemiluminescence biosensing of aflatoxin M1 based on luminol-functionalized, silver nanoparticle-decorated graphene oxide.

A sensitive electrochemiluminescence (ECL) aptasensor for aflatoxin M1 (AFM1) detection by a closed bipolar electrode (BPE) array has been introduced. The thiolated AFM1 aptamer was immobilized on gold nanoparticle-coated magnetic Fe3O4 nanoparticles (Apt-GMNPs). Luminol-functionalized silver nanoparticle-decorated graphene oxide (GO-L-AgNPs) participates in π-π interactions with the unpaired bases of the immobilized aptamer (Apt-GMNPs-GO-L-AgNPs). After the Apt-GMNPs-GO-L-AgNPs were introduced to a gold anodic BPE array, the individual electrodes were subjected to different concentrations of AFM1. Upon the interaction of AFM1 with the aptamers, the GO-L-AgNPs detach from the aptamer; the resulting ECL of luminol and H2O2 at the anodic poles is monitored using a photomultiplier tube (PMT) or smartphone, and the images are analyzed using ImageJ software. This process triggers thionine reduction at the cathodic poles. Under the optimal conditions obtained by a face-centered central composite design (FCCD), the PMT-based detection of the BPE-ECL aptasensor exhibit a linear response over a wide dynamic range from 5 to 150ngmL-1, with a detection limit of 0.01ngmL-1. Additionally, smartphone-based detection shows a linear relationship between the ECL image gray value and the logarithmic concentration of the AFM1 target over a range of 10-200ngmL-1, with a detection limit of 0.05ngmL-1. Furthermore, the BPE-ECL aptasensor was successfully used to detect AFM1 in milk complex media without any serious interferences with reliable reproducibility (average relative standard deviation (RSD = 2.3%)). This smartphone-based detection opens a new horizon for bioanalysis that does not require a trained technician to operate and is a promising technology for point-of-care testing.

Amplified detection of leukemia cancer cells using an aptamer-conjugated gold-coated magnetic nanoparticles on a nitrogen-doped graphene modified electrode.

The increasing demands for early, accurate and ultrasensitive diagnosis of cancers demonstrate the importance of the development of new amplification strategies or diagnostic technologies. In the present study, an aptamer-based electrochemical biosensor for ultrasensitive and selective detection of leukemia cancer cells has been introduced. The thiolated sgc8c aptamer was immobilized on gold nanoparticles-coated magnetic Fe3O4 nanoparticles (Apt-GMNPs). Ethidium bromide (EB), intercalated into the stem of the aptamer hairpin, provides the read-out signal for the quantification of the leukemia cancer cells. After introduction of the leukemia cancer cells onto the Apt-GMNPs, the hairpin structure of the aptamer is disrupted and the intercalator molecules are released, resulting in a decrease of the electrochemical signal. The immobilization of nitrogen-doped graphene nanosheets on the electrode surface provides an excellent platform for amplifying the read-out signal. Under optimal conditions, the aptasensor exhibits a linear response over a wide dynamic range of leukemia cancer cells from 10 to 1×106cellmL-1. The present protocol provides a highly sensitive, selective, simple, and robust method for early stage detection of leukemia cancer. Furthermore, the fabricated aptasensor was successfully used for the detection of leukemia cancer cells in complex media such as human blood plasma, without any serious interference.

Aptamer-conjugated Magnetic Nanoparticles as Targeted Magnetic Resonance Imaging Contrast Agent for Breast Cancer.

Early detection of breast cancer is the most effective way to improve the survival rate in women. Magnetic resonance imaging (MRI) offers high spatial resolution and good anatomic details, and its lower sensitivity can be improved by using targeted molecular imaging. In this study, AS1411 aptamer was conjugated to Fe3O4@Au nanoparticles for specific targeting of mouse mammary carcinoma (4T1) cells that overexpress nucleolin. In vitro cytotoxicity of aptamer-conjugated nanoparticles was assessed on 4T1 and HFFF-PI6 (control) cells. The ability of the synthesized nanoprobe to target specifically the nucleolin overexpressed cells was assessed with the MRI technique. Results show that the synthesized nanoprobe produced strongly darkened T2-weighted magnetic resonance (MR) images with 4T1 cells, whereas the MR images of HFFF-PI6 cells incubated with the nanoprobe are brighter, showing small changes compared to water. The results demonstrate that in a Fe concentration of 45 µg/mL, the nanoprobe reduced by 90% MR image intensity in 4T1 cells compared with the 27% reduction in HFFF-PI6 cells. Analysis of MR signal intensity showed statistically significant signal intensity difference between 4T1 and HFFF-PI6 cells treated with the nanoprobe. MRI experiments demonstrate the high potential of the synthesized nanoprobe as a specific MRI contrast agent for detection of nucleolin-expressing breast cancer cells.

Wireless electrochemiluminescence bipolar electrode array for visualized genotyping of single nucleotide polymorphism.

The development of simple, inexpensive, hand-held, user-friendly biosensor for high throughput and multiplexed genotyping of various single nucleotide polymorphisms (SNPs) in a single run experiment by a nonspecialist user is the main challenge in the analysis of DNA. Visualizing the signal and possibility to monitor SNPs by a digital camera opens a new horizon for the routine applications. In the present manuscript, a novel wireless electrochemiluminescence (ECL) DNA array is introduced for the visualized genotyping of different SNPs on the basis of ECL of luminol/hydrogen peroxide system on a bipolar electrode (BPE) array platform. After modification of anodic poles of the array with the DNA probe and its hybridization with the targets, genotyping of various SNPs is carried out by exposing the array to different monobase modified luminol-platinum nanoparticles (M-L-PtNPs). Upon the hybridization of M-L-PtNPs to mismatch sites, the ECL of luminol is followed using a photomultiplier tube (PMT) or digital camera and the images are analyzed by ImageJ software. This biosensor can detect even thermodynamically stable SNP (G-T mismatches) in the range of 2-600 pM. Also, by combining the advantages of BPE and the high visual sensitivity of ECL, it could be easily expected to achieve sensitive screening of different SNPs. The present biosensor demonstrates the capability for the discrimination between PCR products of normal, heterozygous, and homozygous beta thalassemia genetic disorders.

Carbon nanotube composite coated platinum electrode for detection of Ga(III).

This study demonstrates the application of composite multi-walled carbon nanotube (MWCNT) polyvinylchloride (MWNT-PVC) based on 7-(2-hydroxy-5-methoxybenzyl)-5,6,7,8,9,10-hexahydro-2H benzo [b][1,4,7,10,13] dioxa triaza cyclopentadecine-3,11(4H,12H)-dione ionophore for gallium sensor. The sensor shows a good Nernstian slope of 19.68 ± 0.40 mV/decade in a wide linear range concentration of 7.9 × 10(-7) to 3.2 × 10(-2)M of Ga(NO(3))(3). The detection limit of this electrode is 5.2 × 10(-7)M of Ga(NO(3))(3). This proposed sensor is applicable in a pH range of 2.7-5.0. It has a short response time of about 10s and has a good selectivity over nineteen various metal ions. The practical analytical utility of this electrode is demonstrated by measurement of Ga(III) in river water.