Ultrasensitive aptamer-based on-off assay for lysozyme using a glassy carbon electrode modified with gold nanoparticles and electrochemically reduced graphene oxide

The authors describe an electrochemical aptamer based assay for the determination of the serine protease lysozyme in very low (pM) concentrations. The method is based on the formation of a complex between anti-lysozyme aptamer fragments and lysozyme, and on electrochemical detection by differential pulse voltammetry (DPV) and electrochemical impedance spectroscopy (EIS). The surface of a glassy carbon electrode was modified with a nanocomposite consisting of gold nanoparticles and electrochemically reduced graphene oxide nanosheets (AuNPs/erGO), and the thiolated aptamer was then linked to the AuNPs by self-assembly through Au-S bonds. The interaction of immobilized aptamers with lysozyme leads to the decreased peak current in DPV and increased charge transfer resistance (R_ct) in EIS when using hexacyanoferrate or Methylene Blue as a redox probe. The calibration plot, when applying EIS and working at a typical voltage of ?0.22 V (vs. SCE), is linear over 1.0 to 104.3 pM concentration range, with a detection limit of 0.06 pM (at a signal-to-noise ratio of 3). The respective data for DPV are a 9.6–205.5 pM linear range with a detection limit of 0.24 pM. Depending on the redox marker applied, the method works in the “signal-off” or “signal-on” mode in DPV and EIS protocols, respectively. The sensing interface is high specific for lysozyme and not affected by other proteins. The method was applied to the determination of lysozyme in spiked diluted human serum, and the results agreed well with data obtained with a standard ELISA.

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Graphical abstract The surface of a glassy carbon electrode was modified with a nanocomposite consisting of gold nanoparticles and electrochemically reduced graphene oxide nanosheets (AuNPs/erGO). Then, the thiolated aptamer was linked to the AuNPs by self-assembly through Au-S bonds. The modified electrode was applied to the determination of lysozyme with “signal off” and “signal on” strategies.

     

Voltammetric dopamine sensor based on a gold electrode modified with reduced graphene oxide and Mn3O4 on gold nanoparticles

Microchimica Acta - The authors describe an amperometric sensor for dopamine (DA) which has a working potential as low as +0.02 V (vs. SCE). It makes use of a nanocomposite consisting of...     

Amperometric detection of nitric oxide using a glassy carbon electrode modified with gold nanoparticles incorporated into a nanohybrid composed of reduced graphene oxide and Nafion

Microchimica Acta - The authors show that the electrocatalytic performance toward the detection of nitric oxide (NO) can be enhanced by making use of gold nanoparticles (AuNP) in a matrix...     

Impedance immunosensor for bovine interleukin-4 using an electrode modified with reduced graphene oxide and chitosan

Microchimica Acta - We have developed an impedimetric immunosensor for the determination of bovine interleukin-4, a cytokine involved in the immune response to certain parasites and the development...     

Electrochemical detection of selenium using glassy carbon electrode modified with reduced graphene oxide

In this work, a simple experimental procedure was reported for the electroanalytical determination of selenium (IV) using reduced graphene oxide (rGO) to modify glassy carbon electrode (GCE). The rGO was obtained by reduction of graphene oxide obtained via Hummer’s method. The synthesised rGO was characterised using X-ray diffraction, Raman spectroscopy, scanning electron microscope (SEM), energy-dispersive spectroscopy and transmission Electron microscopy (TEM). GCE was modified with rGO and the electrochemical properties of the bare and modified electrode were investigated using cyclic voltammetry and electrochemical impedance spectroscopy. The results obtained showed that the modified electrode exhibited more excellent electrochemical properties than the bare GCE. The optimum conditions for detection of selenium in water using square wave anodic stripping voltammetry were as follows: deposition potential ?500 mV, pH 1, pre-concentration time of 240 s and 0.1 M nitric acid was used as supporting electrolyte. The linear regression equation obtained was I (µA) = 0.8432C + 9.2359 and the detection limit was calculated to be 0.85 μg L^?1. However, Cu(II) and Cd(II) are the two cations that interfered in the analysis of selenium in water.

The sensor was also applied for real sample water analysis and the result obtained was affirmed with inductively coupled plasma optical emission spectroscopic method. It is believed that our proposed sensor hold promise for practical application.     

Sandwich electrochemical thrombin assay using a glassy carbon electrode modified with nitrogen- and sulfur-doped graphene oxide and gold nanoparticles

Graphene oxide doped with nitrogen and sulfur was decorated with gold nanoparticles (AuNP-SN-GO) and applied as a substrate to modify a glassy carbon electrode (GCE). An aptamer against the model protein thrombin was self-assembled on the modified GCE which then was exposed to thrombin. Following aptamer-thrombin interaction, biotin-labeled DNA and aptamer 2 are immobilized on another AuNP-SN-GO hybrid and then are reacted with the thrombin/AuNP-SN-GO/GCE to form a sandwich. The enzyme label horseradish peroxidase (HRP) was then attached to the electrode by biotin–avidin interaction. HRP catalyzes the oxidation of hydroquinone by hydrogen peroxide. This generates a strong electrochemical signal that increases linearly with the logarithm of thrombin concentration in the range from 1.0?×?10^?13 M to 1.0?×?10^?8 M with a detection limit of 2.5?×?10^?14 M (S/N?=?3). The assay is highly selective. It provides a promising strategy for signal amplification. In our perception, it has a large potential for sensitive and selective detection of analytes for which appropriate aptamers are available.

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Graphic abstract A sandwich-type electrochemical aptasensor is fabricated for detection of thrombin using a glassy carbon electrode modified with nitrogen- and sulfur-doped graphene oxide and gold nanoparticles.

     

Determination of trypsin activity using a gold electrode modified with a nanocover composed of graphene oxide and thionine

We report on an electrochemical method for the determination of the activity of trypsin. A multi-functional substrate peptide (HHHAKSSATGGC-HS) is designed and immobilized on a gold electrode. The three His residues in the N-terminal are able to recruit thionine-loaded graphene oxide (GO/thionine), a nanocover adopted for signal amplification. Once the peptide is cleaved under enzymatic catalysis by trypsin (cleavage site: Lys residue), the His residues leave the electrode, and the GO/thionine cannot cover the peptide-modified electrode anymore. Thus, the changes of the electrochemical signal of thionine, typically acquired at a voltage of -0.35 V, can be used to determine the activity of trypsin. A detection range of 1 × 10⁻⁴ to 1 U, with a detection limit of 3.3 × 10⁻⁵ U, can be achieved, which is better than some currently available methods. In addition, the method is highly specific, facile, and has the potential for the detection of trypsin-like proteases.

Graphene oxide was adopted as a nanocover for the development of a sensitive electrochemical method to detect the activity of trypsin.

     

Photoelectrochemical determination of hydrogen peroxide using a gold electrode modified with fluorescent gold nanoclusters and graphene oxide

Nanocomposites consisting of gold nanoclusters and graphene oxide (AuNC/GO) were prepared and investigated with respect to the design of new sensors for hydrogen peroxide (H₂O₂). The AuNC/GO hybrid nanomaterials were deposited on a gold electrode by the layer-by-layer assembly method, where they showed enhanced photoelectrical and sensing properties. The presence of graphene oxide improves the photoinduced electron separation efficiency of the AuNCs, as well as the catalytic effect of AuNCs on the electroreduction of H₂O₂. Compared to an electrode modified with AuNCs only, the new electrodes display a more than ten-fold enhanced photocurrent at a working voltage of -500 mV (vs. Ag/AgCl), higher sensitivity for H₂O₂ (25.76 nA?mM^?1), lower LOD (2 μM) and extended linear range (from 30 μM to 5 mM). The sensors were applied to the determination of H₂O₂ extracted from living human umbilical vein endothelial cells stimulated by angiotensin II.

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Graphical abstract Graphene oxide (GO) not only improves the photoinduced charge separation efficiency of fluorescent gold nanoclusters (AuNCs) based photoelectrochemical sensors, but also enhances the catalytic property of AuNCs on the detection of hydrogen peroxide (H₂O₂).

     

Amperometric enzymatic determination of bisphenol A using an ITO electrode modified with reduced graphene oxide and Mn3O4 nanoparticles in a chitosan matrix

Microchimica Acta - The authors describe a nanostructured hybrid platform for amperometic determination of the plasticizer bisphenol A (BPA). It consists of two dimensional sheets of reduced...     

Dopamine sensor based on a glassy carbon electrode modified with a reduced graphene oxide and palladium nanoparticles composite

We report on a sensitive electrochemical sensor for dopamine (DA) based on a glassy carbon electrode that was modified with a nanocomposite containing electrochemically reduced graphene oxide (RGO) and palladium nanoparticles (Pd-NPs). The composite was characterized by scanning electron microscopy, energy dispersive spectroscopy, and electrochemical impendence spectroscopy. The electrode can oxidize DA at lower potential (234 mV vs Ag/AgCl) than electrodes modified with RGO or Pd-NPs only. The response of the sensor to DA is linear in the 1–150 μM concentration range, and the detection limit is 0.233 μM. The sensor was applied to the determination of DA in commercial DA injection solutions.

Voltammetric determination of nitric oxide using a glassy carbon electrode modified with a nanohybrid consisting of myoglobin,gold nanorods,and reduced graphene oxide

Microchimica Acta - Myoglobin-modified gold nanorods incorporating reduced graphene oxide (rGO) were fabricated and deposited on a glassy carbon electrode (GCE) to obtain a sensor for nitric oxide...     

Label-free immunosensing of microcystin-LR using a gold electrode modified with gold nanoparticles

The hepatotoxic microcystins, especially microcystin?CLR (MC?CLR), are causing serious problems to public health and fisheries. We describe here a label-free amperometric immunosensor for rapid determination of MC?CLR in water sample. The sensor was prepared by immobilizing antibody on a gold electrode coated with L-cysteine-modified gold nanoparticles. The stepwise self-assembly of the immunosensor was monitored and characterized by means of electrochemical impedance spectroscopy and differential pulse voltammetry. A 0.60?mmol L^?1 solution of hydroquinone was used as the electron mediator. The immunosensor was incubated with MC?CLR at 25?°C for 20?min, upon which the differential pulse voltammetric current changed linearly over the concentration range from 0.05 to 15.00???g L^?1, with a detection limit of 20?ng L^?1. The developed biosensor was used to determine MC?CLR in spiked crude algae samples. The recovery was in the range from 95.6 to 105%. This method is simple, economical and efficient, this making it potentially suitable for field analysis of MC-LR in crude algae and water samples.

Determination of hydroxylamine using a carbon paste electrode modified with graphene oxide nano sheets

A carbon paste electrode that was chemically modified with 3-(4'-amino-3'-hydroxy-biphenyl-4-yl)-acrylic acid (3,4-AA) was used as a selective electrochemical sensor for the detection of hydroxylamine. Cyclic voltammetry (CV), choronoamperometry (CHA) and square wave voltammetry (SWV) were used to investigate oxidation of hydroxylamine in aqueous solution. Under optimized concentration the electrocatalytic oxidation current peak for hydroxylamine increased linearly with concentration in the range of 0.025–10.0 μM. The detection limits for hydroxylamine was 0.012 μM. Finally, the modified electrode was applied to detection hydroxylamine in water samples.     

Amperometric determination of L-cysteine using a glassy carbon electrode modified with palladium nanoparticles grown on reduced graphene oxide in a Nafion matrix

An amperometric sensor for L-Cys is described which consists of a glassy carbon electrode (GCE) that was modified with reduced graphene oxide placed in a Nafion film and decorated with palladium nanoparticles (PdNPs). The film was synthesized by a hydrothermal method. The PdNPs have an average diameter of about 10 nm and a spherical shape. The modified GCE gives a linear electro-oxidative response to L-Cys (typically at +0.6 V vs. SCE) within the 0.5 to 10 μM concentration range. Other figures of merit include a response time of less than 2 s, a 0.15 μM lower detection limit (at signal to noise ratio of 3), and an analytical sensitivity of 1.30 μA·μM^?1·cm^?2. The sensor displays selectivity over ascorbic acid, uric acid, dopamine, hydrogen peroxide, urea, and glucose. The modified GCE was applied to the determination of L-Cys in human urine samples and gave excellent recoveries.

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Graphical abstract Spherical palladium nanoparticles (PdNPs) on reduced graphene oxide-Nafion (rGO-Nf) films were synthesized using a hydrothermal method. This nanohybrid was used for modifying a glassy carbon electrode to develop a sensor electrode for detecting L-cysteine that has fast response (less than 2 s), low detection limit (0.15 μM), and good sensitivity (0.092 μA μM^-1 cm^-2).