An alternative ready-to-use electrochemical immunosensor for point-of-care COVID-19 diagnosis using graphene screen-printed electrodes coupled with a 3D-printed portable potentiostat.

A severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a cause of worldwide Coronavirus 2019 (COVID-19) disease pandemic. It is thus important to develop ultra-sensitive, rapid and easy-to-use methods for the identification of COVID-19 infected patients. Herein, an alternative electrochemical immunosensor based on poly(pyrrolepropionic acid) (pPPA) modified graphene screen-printed electrode (GSPE) was proposed for rapid COVID-19 detection. The method was based on a competitive enzyme immunoassay process utilizing horseradish peroxidase (HRP)-conjugated SARS-CoV-2 as a reporter binding molecule to compete binding with antibody against the SARS-CoV-2 receptor binding domain (SARS-CoV-2 RBD) protein. This strategy enhanced the current signal via the enzymatic reaction of HRP-conjugated SARS-CoV-2 RBD antibody on the electrode surface. The modification, immobilization, blocking, and detection processes were optimized and evaluated by amperometry. The quantitative analysis of SARS-CoV-2 was conducted based on competitive enzyme immunoassay with amperometric detection using a 3D-printed portable potentiostat for point-of-care COVID-19 diagnosis. The current measurements at -0.2 V yielded a calibration curve with a linear range of 0.01-1500 ng mL-1 (r2 = 0.983), a low detection limit of 2 pg mL-1 and a low quantification limit of 10 pg mL-1. In addition, the analyzed results of practical samples using the developed method were successfully verified with ELISA and RT-PCR. Therefore, the proposed portable electrochemical immunosensor is highly sensitive, rapid, and reliable. Thus, it is an alternative ready-to-use sensor for COVID-19 point-of-care diagnosis.

A facile one-pot synthesis of magnetic iron oxide nanoparticles embed N-doped graphene modified magnetic screen printed electrode for electrochemical sensing of chloramphenicol and diethylstilbestrol.

Trace determination of antibacterial agents is crucial to minimize risks of human intoxication and in the prevention of serious environmental impacts. Herein, a simple one-pot solvothermal synthesis approach for a magnetic iron oxide embed nitrogen-doped graphene (MIO@NG) nanohybrid was fabricated without the addition of any extra reductant and its application towards ultrasensitive chloramphenicol (CAP) and diethylstilbestrol (DES) electrochemical sensor is demonstrated to screen for antibiotic residue contamination in milk samples. The prepared nanohybrid was modified on a magnetic screen-printed electrode (MSPE) to make it portable for on-site detection. The determination of two additive drugs, CAP and DES, was achieved based on the reduction current response at MIO@NG modified MSPE (MIO@NG/MSPE) to eliminate interference as far as possible. Uniform dispersed MIO nanoparticles are grown in situ on the surface of nitrogen-doped graphene sheets. The morphology of MIO@NG was confirmed by transmission electron microscopy (TEM) analysis. The chemical structure of the prepared MIO@NG was characterized by x-ray diffraction (XRD), x-ray photoemission spectroscopy (XPS), Raman spectroscopy, and extended x-ray absorption fine structure (EXAFS). Moreover, the superparamagnism property was investigated by vibrating sample magnetometry (VSM). The electrochemical properties of MIO@NG were evaluated with cyclic voltammetry (CV) and square wave voltammetry (SWV). Sensor performance was evaluated by testing the electrochemical activity of CAP and DES in the presence of interferences. The MIO@NG modified electrode presented superior electrochemical performance, including high sensitivity, high catalytic activity, ultimate sensitivity, very fast detection, selectivity, and excellent performance. The MIO@NG modified electrode demonstrated a detection limit of 10 nM for the detection of CAP and 6.5 nM for DES with satisfactory recovery in real samples.

Magnetic Nanoparticle-Reduced Graphene Oxide Nanocomposite as a Novel Bioelectrode for Mediatorless-Membraneless Glucose Enzymatic Biofuel Cells.

In this work, an enzymatic biofuel cell (EBC) based on a membraneless and mediatorless glucose enzymatic fuel cell system was constructed for operation in physiological conditions (pH 7.0 and temperature 37 °C). The new platform EBC made of nanocomposite, including magnetic nanoparticles (Fe3O4 NPs) and reduced graphene oxide (RGO), was used for the immobilization of glucose oxidase (GOD) as bioanode and bilirubin oxidase (BOD) as biocathode. The EBC bioelectrodes were fabricated without binder or adhesive agents for immobilized enzyme and the first EBC using superparamagnetic properties with Fe3O4 NPs has been reported. The performance of the EBC was evaluated with promising results. In EBC tests, the maximum power density of the EBC was 73.7 µW cm-2 and an open circuit voltage (OCV) as +0.63 V with 5 mM of glucose concentration for the physiological condition of humans. The Fe3O4-RGO nanocomposite offers remarkable enhancement in large surface areas, is a favorable environment for enzyme immobilization, and facilitates electron transfer between enzymes and electrode surfaces. Fe3O4 and RGO have been implied as new promising composite nanomaterials for immobilizing enzymes and efficient platforms due to their superparamagnetism properties. Thus, glucose EBCs could potentially be used as self-powered biosensors or electric power sources for biomedical device applications.

Self-assembly of glucose oxidase on reduced graphene oxide-magnetic nanoparticles nanocomposite-based direct electrochemistry for reagentless glucose biosensor.

A novel approach of the immobilization of a highly selective and stable glucose biosensor based on direct electrochemistry was fabricated by a self-assembly of glucose oxidase (GOD) on reduced graphene oxide (RGO) covalently conjugated to magnetic nanoparticles (Fe3O4 NPs) modified on a magnetic screen-printed electrode (MSPE). The RGO-Fe3O4 nanocomposite has remarkable enhancement in large surface areas, is favorable environment for enzyme immobilization, facilitates electron transfer between enzymes and electrode surfaces and possesses superparamagnetism property. The morphology and electrochemical properties of RGO-Fe3O4/GOD were characterized by scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FTIR), Raman spectroscopy, cyclic voltammetry (CV) and amperometry. The modified electrode was a fast, direct electron transfer with an apparent electron transfer rate constant (ks) of 13.78s-1. The proposed biosensor showed fast amperometric response (3s) to glucose with a wide linear range from 0.05 to 1mM, a low detection limit of 0.1µM at a signal to noise ratio of 3 (S/N=3) and good sensitivity (5.9µA/mM). The resulting biosensor has high stability, good reproducibility, excellent selectivity and successfully applied detection potential at -0.45V. This mediatorless glucose sensing used the advantages of covalent bonding and self-assembly as a new approach for immobilizing enzymes without any binder. It would be worth noting that it opens a new avenue for fabricating excellent electrochemical biosensors. NOVELTY STATEMENT: This is a new approach that reporting the immobilization of glucose oxidase on reduced graphene oxide (RGO) covalently conjugated to magnetic nanoparticles (Fe3O4 NPs) by electrostatic interaction and modified screen printed electrode. We propose the reagentless with fabrication method without binder and adhesive agents for immobilized enzyme. Fe3O4 NPs increasing surface area to enhance the immobilization and prevent the leaching of enzymes at electrode surfaces by magnetic stickers which is improve the stability of the biosensor. Based on this synthesis technique, it is a good new strategy and simple used to fabrication of third-generation glucose biosensor and this nanocomposite could be used as a platform for disposable biosensor and biofuel cell applications.

Highly cytocompatible and flexible three-dimensional graphene/polydimethylsiloxane composite for culture and electrochemical detection of L929 fibroblast cells.

Recently, three-dimensional graphene interconnected network has attracted great interest as a scaffold structure for tissue engineering due to its high biocompatibility, high electrical conductivity, high specific surface area and high porosity. However, free-standing three-dimensional graphene exhibits poor flexibility and stability due to ease of disintegration during processing. In this work, three-dimensional graphene is composited with polydimethylsiloxane to improve the structural flexibility and stability by a new simple two-step process comprising dip coating of polydimethylsiloxane on chemical vapor deposited graphene/Ni foam and wet etching of nickel foam. Structural characterizations confirmed an interconnected three-dimensional multi-layer graphene structure with thin polydimethylsiloxane scaffold. The composite was employed as a substrate for culture of L929 fibroblast cells and its cytocompatibility was evaluated by cell viability (Alamar blue assay), reactive oxygen species production and vinculin immunofluorescence imaging. The result revealed that cell viability on three-dimensional graphene/polydimethylsiloxane composite increased with increasing culture time and was slightly different from a polystyrene substrate (control). Moreover, cells cultured on three-dimensional graphene/polydimethylsiloxane composite generated less ROS than the control at culture times of 3-6 h. The results of immunofluorescence staining demonstrated that fibroblast cells expressed adhesion protein (vinculin) and adhered well on three-dimensional graphene/polydimethylsiloxane surface. Good cell adhesion could be attributed to suitable surface properties of three-dimensional graphene/polydimethylsiloxane with moderate contact angle and small negative zeta potential in culture solution. The results of electrochemical study by cyclic voltammetry showed that an oxidation current signal with no apparent peak was induced by fibroblast cells and the oxidation current at an oxidation potential of +0.9 V increased linearly with increasing cell number. Therefore, the three-dimensional graphene/polydimethylsiloxane composite exhibits high cytocompatibility and can potentially be used as a conductive substrate for cell-based electrochemical sensing.