Reduced Graphene Oxide/Organic Dye Composites for Bioelectroconversion of Saccharides: Application for Detection of Saccharides and α-Amylase Assessments.

In this study, PQQ-dependent glucose dehydrogenase (PQQ-GDH) was immobilized onto reduced graphene oxide (rGO) modified with organic dyes from three different classes (acridine, arylmethane, and diazo); namely, neutral red (NR), malachite green (MG), and congo red (CR) formed three types of biosensors. All three rGO/organic dye composites were characterized by scanning electron microscopy, X-ray photoelectron spectroscopy, and Raman spectroscopy. The impact of three rGO/organic dye modifications employed in bioelectrocatalytic systems on changes in enzyme activity and substrate selectivity was investigated. The highest sensitivity of 39 µA/cm2 was obtained for 1 mM of glucose when a rGO_MG/PQQ-GDH biosensor was used. A significant improvement in the electrochemical response of biosensors was attributed to the higher amount of pyrrolic nitrogen groups on the surface of the rGO/organic dye composites. Modifications of rGO by NR and MG not only improved the surfaces for efficient direct electron transfer (DET) but also influenced the enzyme selectivity through proper binding and orientation of the enzyme. The accuracy of the biosensor's action was confirmed by the spectrophotometric analysis. Perspectives for using the proposed bioelectrocatalytic systems operating on DET principles for total or single monosaccharide and/or disaccharide determination/bioconversion systems or for diagnoses have been presented through examples of bioconversion of D-glucose, D-xylose, and maltose.

Capacitance-Based Biosensor for the Measurement of Total Loss of L-Amino Acids in Human Serum during Hemodialysis.

In this paper, we present a biosensor based on a gold nanoparticle (AuNP)-modified Pt electrode with an adjusted membrane containing cross-linked L-amino acid oxidase for the detection and quantification of total L-amino acids. The designed biosensor was tested and characterized using the capacitance-based principle, capacitance measurements after electrode polarization, disconnection from the circuit, and addition of the respective amount of the analyte. The method was implemented using the capacitive and catalytic properties of the Pt/AuNP electrode; nanostructures were able to store electric charge while at the same time catalyzing the oxidation of the redox reaction intermediate H2O2. In this way, the Pt/AuNP layer was charged after the addition of analytes, allowing for much more accurate measurements for samples with low amino acid concentrations. The combined biosensor electrode with the capacitance-based measurement method resulted in high sensitivity and a low limit of detection (LOD) for hydrogen peroxide (4.15 µC/µM and 0.86 µM, respectively) and high sensitivity, a low LOD, and a wide linear range for L-amino acids (0.73 µC/µM, 5.5 µM and 25-1500 µM, respectively). The designed biosensor was applied to measure the relative loss of amino acids in patients undergoing renal replacement therapy by analyzing amino acid levels in diluted serum samples before and after entering/leaving the hemodialysis apparatus. In general, the designed biosensor in conjunction with the proposed capacitance-based method was clinically tested and could also be applied for the detection of other analytes using analyte-specific oxidases.

Technical suitability and reliability of an in vivo and non-invasive biosensor-type glucose assessment as a potential biomarker for multiple stressors in fishes: an evaluation on Salmonids.

Regardless of the wide use of glucose measurements in stress evaluation, there are some inconsistencies in its acceptance as a stress marker. To meet the challenge and test the reliability/suitability of glucose measurement in practice, we simulated different environmental/anthropogenic exposure scenarios in this study. We aimed to provoke stress in fish followed by a 2-week stress recovery period and under the cumulative effect of leachate fish exposed to pathogenic oomycetes (Saprolegnia parasitica) to represent a possible infection in fish. We selected stream-resident and anadromous brown trout ecotypes (Salmo trutta) representing salmonids with different migratory behaviour strategies. Here, we analysed glucose content in fish-holding water, blood and gills to determine glucose suitability as a potential biomarker of fish response to environmental challenges. Additionally, swimming behavioural parameters and haematocrit were measured. The results indicated that the quantity of glucose released in the holding water of stressed fish increased considerably and remained substantially higher throughout the stress recovery period than the control level. Correspondingly, the circulating levels of glucose in blood and gills decreased over time in fish exposed to different stressors. A significant decrease in swimming activity of fish was observed during the first hours of leachate exposure and increased in fish exposed to S. parasitica compared to control. Our study is the first to ensure the validity and reliability of glucose response in evaluating physiological stress in fish under chemical and biological stimuli, indicating its sensitivity and response range of glucose measurement in fish-holding water.

Reagentless D-Tagatose Biosensors Based on the Oriented Immobilization of Fructose Dehydrogenase onto Coated Gold Nanoparticles- or Reduced Graphene Oxide-Modified Surfaces: Application in a Prototype Bioreactor.

As electrode nanomaterials, thermally reduced graphene oxide (TRGO) and modified gold nanoparticles (AuNPs) were used to design bioelectrocatalytic systems for reliable D-tagatose monitoring in a long-acting bioreactor where the valuable sweetener D-tagatose was enzymatically produced from a dairy by-product D-galactose. For this goal D-fructose dehydrogenase (FDH) from Gluconobacter industrius immobilized on these electrode nanomaterials by forming three amperometric biosensors: AuNPs coated with 4-mercaptobenzoic acid (AuNP/4-MBA/FDH) or AuNPs coated with 4-aminothiophenol (AuNP/PATP/FDH) monolayer, and a layer of TRGO on graphite (TRGO/FDH) were created. The immobilized FDH due to changes in conformation and spatial orientation onto proposed electrode surfaces catalyzes a direct D-tagatose oxidation reaction. The highest sensitivity for D-tagatose of 0.03 ± 0.002 µA mM-1cm-2 was achieved using TRGO/FDH. The TRGO/FDH was applied in a prototype bioreactor for the quantitative evaluation of bioconversion of D-galactose into D-tagatose by L-arabinose isomerase. The correlation coefficient between two independent analyses of the bioconversion mixture: spectrophotometric and by the biosensor was 0.9974. The investigation of selectivity showed that the biosensor was not active towards D-galactose as a substrate. Operational stability of the biosensor indicated that detection of D-tagatose could be performed during six hours without loss of sensitivity.

The Synergy of Thermally Reduced Graphene Oxide in Amperometric Urea Biosensor: Application for Medical Technologies.

Thermally reduced graphene oxide (TRGO) is a graphene-based nanomaterial that has been identified as promising for the development of amperometric biosensors. Urease, in combination with TRGO, allowed us to create a mediator-free amperometric biosensor with the intention of precise detection of urea in clinical trials. Beyond simplicity of the technology, the biosensor exhibited high sensitivity (2.3 ± 0.1 µA cm-2 mM-1), great operational and storage stabilities (up to seven months), and appropriate reproducibility (relative standard deviation (RSD) about 2%). The analytical recovery of the TRGO-based biosensor in urine of 101 ÷ 104% with RSD of 1.2 ÷ 1.7% and in blood of 92.7 ÷ 96.4%, RSD of 1.0 ÷ 2.5%, confirmed that the biosensor is acceptable and reliable. These properties allowed us to apply the biosensor in the monitoring of urea levels in samples of urine, blood, and spent dialysate collected during hemodialysis. Accuracy of the biosensor was validated by good correlation (R = 0.9898 and R = 0.9982) for dialysate and blood, utilizing approved methods. The advantages of the proposed biosensing technology could benefit the development of point-of-care and non-invasive medical instruments.

Fine structure and related properties of the assembleable carbon nanotubes based electrode for new family of biosensors with chooseable selectivity.

Surfaces of constituent parts of biosensors based on single wall carbon nanotube layer were investigated and compare for properly functioning and faulty biosensors. Though the original technology is acceptable for changing of the selectivity, only glucose sensitive biosensors are investigated. Based on the results of the study, a correlation between the features of the nanoscale structures and parameters of amperiometric biosensors for assemblage of which an innovative approach is described. Original template of the electrodes has been prepared on a base of single wall carbon nanotube layer deposited on the supporting polycarbonate membrane. Original immobilisation of enzymes within special membrane allows functional modification of biosensors being accomplished by simple replacement of the enzymatic membrane. The original technology leads to a novel family of biosensors acceptable for detection of wide range of carbohydrates. The morphology and the local electric properties of the constituent parts of the biosensors are characterized by scanning probe microscopy. The sensitivity, selectivity and stability are described for typical types of the biosensors.

Application of oxygen-independent biosensor for testing yeast fermentation capacity.

The pyrroloquinoline quinone (PQQ)-dependent soluble glucose dehydrogenase based carbon paste electrodes were investigated and applied for glucose monitoring in the oxygen deficient media. Reagentless biosensors possessing a wide linear range (up to 5 mM glucose with a detection limit of 0.12 mM) were designed. The oxygen-insensitive response of the biosensor creates the opportunity to use it as a flow-through device for continuous monitoring of glucose in media during the wine yeast fermentation process. The analysis of glucose assimilation rate by yeast strains using the developed biosensor correlated well (R2=0.9938) with convenient yeast testing methods.