Search

The improved binding ability of graphene–nanoparticle composites to proteins or molecules can be utilized to develop new cell-based assays. In this study, we fabricated reduced graphene oxide–gold nanoparticles (rGO-AuNP) electrodeposited onto a transparent indium tin oxide (ITO) electrode and investigated the feasibility of the electrochemical impedance monitoring of cell growth. The electrodeposition of rGO–AuNP on the ITO was optically and electrochemically characterized in comparison to bare, rGO-, and AuNP-deposited electrodes. The cell growth on the rGO–AuNP/ITO electrode was analyzed via electrochemical impedance measurement together with the microscopic observation of HEK293 cells transfected with a green fluorescent protein expression vector. The results showed that rGO–AuNP was biocompatible and induced an increase in cell adherence to the electrode when compared to the bare, AuNP-, or rGO-deposited ITO electrode. At 54 h cultivation, the average and standard deviation of the saturated normalized impedance magnitude of the rGO–AuNP/ITO electrode was 3.44 ± 0.16, while the value of the bare, AuNP-, and rGO-deposited ITO electrode was 2.48 ± 0.15, 2.61 ± 0.18, and 3.01 ± 0.25, respectively. The higher saturated value of the cell impedance indicates that the impedimetric cell-based assay has a broader measurement range. Thus, the rGO–AuNP/ITO electrode can be utilized for label-free and real-time impedimetric cell-based assays with wider dynamic range. [ABSTRACT FROM AUTHOR]

The alcohol oxidase (AOx) cDNA from Aspergillus terreus MTCC6324 with an open reading frame (ORF) of 2001 bp was constructed from n-hexadecane induced cells and expressed in Escherichia coli with a yield of ∼4.2 mg protein g−1 wet cell. The deduced amino acid sequences of recombinant rAOx showed maximum structural homology with the chain B of aryl AOx from Pleurotus eryngii. A functionally active AOx was achieved by incubating the apo-AOx with flavin adenine dinucleotide (FAD) for ∼80 h at 16°C and pH 9.0. The isoelectric point and mass of the apo-AOx were found to be 6.5±0.1 and ∼74 kDa, respectively. Circular dichroism data of the rAOx confirmed its ordered structure. Docking studies with an ab-initio protein model demonstrated the presence of a conserved FAD binding domain with an active substrate binding site. The rAOx was specific for aryl alcohols and the order of its substrate preference was 4-methoxybenzyl alcohol >3-methoxybenzyl alcohol>3, 4-dimethoxybenzyl alcohol > benzyl alcohol. A significantly high aggregation to ∼1000 nm (diameter) and catalytic efficiency (kcat/Km) of 7829.5 min−1 mM−1 for 4-methoxybenzyl alcohol was also demonstrated for rAOx. The results infer the novelty of the AOx and its potential biocatalytic application. [ABSTRACT FROM AUTHOR]

Rheumatoid arthritis (RA) is a chronic autoimmune disease that produces a progressive inflammatory response that leads to severe pain, swelling, and stiffness in the joints of hands and feet, followed by irreversible damage of the joints. The authors developed a miniaturized, label-free electrochemical impedimetric immunosensor for the sensitive and direct detection of arthritis Anti-CCP-ab biomarker. An interdigitated-chain-shaped microelectrode array (ICE) was fabricated by taking the advantage of microelectromechanical systems. The fabricated ICE was modified with a self-assembled monolayer (SAM) of Mercaptohexanoic acid (MHA) for immobilization of the synthetic peptide bio-receptor (B-CCP). The B-CCP was attached onto the surface of SAM modified ICE through a strong avidin-biotin bio-recognition system. The modified ICE surface with the SAM and bio-molecules (Avidin, B-CCP, Anti-CCP-ab and BSA) was morphologically and electrochemically characterized. The change in the sensor signal upon analyte binding on the electrode surface was probed through the electrochemical impedance spectroscopy (EIS) property of charge-transfer resistance (Rct) of the modified electrodes. EIS measurements were target specific and the sensor response was linearly increased with step wise increase in target analyte (Anti-CCP-ab) concentrations. The developed sensor showed a linear range for the addition of Anti-CCP-ab between 1 IU mL−1 → 800 IU mL−1 in phosphate buffered saline (PBS) and Human serum (HS), respectively. The sensor showed a limit of detection of 0.60 IU mL−1 and 0.82 IU mL−1 in the PBS and HS, respectively. The develop bio-electrode showed a good reproducibility (relative standard deviation (RSD), 1.52%), selectivity and stability (1.5% lost at the end of 20th day) with an acceptable recovery rate (98.0% → 101.18%) and % RSD's for the detection of Anti-CCP-ab in spiked HS samples. [ABSTRACT FROM AUTHOR]

Individuals with diabetes can benefit considerably from continuous blood glucose monitoring. To address this challenge, a proof-of-concept was performed for continuous glucose monitoring (CGM) based on an enzymeless porous nanomaterial (pNM)-modified microneedle electrode array (MNEA). The pNM sensing layer was electrochemically deposited on MNs by applying a fixed negative current of −2.5 mA cm˗2 for 400 s. The pNM-modified MNEA was packed using a biocompatible Nafion ionomer. The fabricated MNEAs were 600 × 100 × 150 µm in height, width, and thickness, respectively. The surfaces of the modified MNs were characterized by scanning electron microscopy (SEM) and energy dispersive X-ray analysis (EDX), X-ray diffraction (XRD) and X-ray photoelectron spectroscopy (XPS). The fabricated MNEAs showed a wide dynamic range (1–30 mM) in phosphate-buffered saline (PBS) and in artificial interstitial fluid (ISF), with good sensitivities (PBS: 1.792 ± 0.25 µA mM−1 cm−2, ISF: 0.957 ± 0.14 µA mM−1 cm−2) and low detection limits (PBS: 7.2 µM, ISF: 22 µM). The sensor also showed high stability (loss of 3.5% at the end of 16 days), selectivity, and reproducibility (Relative standard deviations (RSD) of 1.64% and 0.70% for intra- and inter-assay, respectively) and a good response time (2 s) with great glucose recovery rates in ISF (98.7–102%). [ABSTRACT FROM AUTHOR]

An rGO-Ag@SiO 2 nanocomposite-based electrochemical sensor was developed to detect etidronic acid (EA) using the differential pulse voltammetric (DPV) technique. Rapid self-assembly of the rGO-Ag@SiO 2 nanocomposite was accomplished through probe sonication. The developed rGO-Ag@SiO 2 nanocomposite was used as an electrochemical sensing platform by drop-casting on a gold (Au) printed circuit board (PCB). Cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS) confirmed the enhanced electrochemical active surface area (ECASA) and low charge transfer resistance (R ct ) of the rGO-Ag@SiO 2 /Au PCB. The accelerated electron transfer and the high number of active sites on the rGO-Ag@SiO 2 /Au PCB resulted in the electrochemical detection of EA through the DPV technique with a limit of detection (LOD) of 0.68 μM and a linear range of 2.0-200.0 μM. The constructed DPV sensor exhibited high selectivity toward EA, high reproducibility in terms of different Au PCBs, excellent repeatability, and long-term stability in storage at room temperature (25 °C). The real-time application of the rGO-Ag@SiO 2 /Au PCB for EA detection was investigated using EA-based pharmaceutical samples. Recovery percentages between 96.2% and 102.9% were obtained. The developed DPV sensor based on an rGO-Ag@SiO 2 /Au PCB could be used to detect other electrochemically active species following optimization under certain conditions.