Bio-Electroanalysis Performance of Heme Redox-Center for π-π Interaction Bonding of a Methylene Blue-Graphene Modified Electrode.

Hemeprotein detection has motivated extensive research on the direct reaction of a heme molecule and a redox dye. The present study used methylene blue as both donor and acceptor for a redox reaction. First, the solid phases of methylene blue (MB) and graphene (GP) formed a π-π interaction bond at the aromatic rings. The conductivity of GP was better than that of carbon in a carbon electrode (CE). Then, the working CE was modified using strong adsorption of MB/GP on the electrode surface. The surface of the electrode was investigated using a modified and an unmodified electrode. The electrode's properties were studied using voltammograms of redox couple K3[Fe(CN)6]3-/4-. Its reaction was used to find the active area of the modified electrode, which was 1.76 times bigger than that of the unmodified electrode. The surface coverage values of the modified and unmodified electrodes were 8.17 × 10-6 and 1.53 × 10-5 mol/cm2, respectively. This research also studied the application of hemeprotein detection. Hemoglobin (Hb), myoglobin (Mb), and cytochrome c (Cyt. C) were studied by the reaction of Fe (III/II) at the heme-redox center. The electrocatalytic reaction between MB/GP and hemeproteins produced an anodic peak at 0.35 V for Hb, Mb, and Cyt. C. This nanohybrid film enhanced electron transfer between protein molecules and the modified carbon electrode. The amperometric measurements show that the limit of detection was 0.2 µM, 0.3 µM, and 0.1 µM for Hb, Mb, and Cyt. C, respectively. The measurement spanned a linear range of 0.2 µM to 5 µM, 0.3 µM to 5 µM, and 0.1 µM to 0.7 µM for Hb, Mb, and Cyt. C, respectively. Hb showed the lowest sensitivity compared with Mb and Cyt. C due to the role of steric hindrance in the hemeprotein specificity structure. This study offers a simple and efficient fabrication platform for electrochemical sensors for hemeproteins. When compared to other complex immobilization processes, the fabrication method for this sensor has many benefits, including no need for special chemicals and easy preparation and electrode modification-both of which are crucial for the development of electrochemical sensing devices.

Salivary Creatinine Detection Using a Cu(I)/Cu(II) Catalyst Layer of a Supercapacitive Hybrid Sensor: A Wireless IoT Device To Monitor Kidney Diseases for Remote Medical Mobility.

The stress-free electrochemical-based sensor equipped with the Internet of Things (IoT) device for salivary creatinine determination was fabricated for point-of-care (POC) diagnosis of advanced kidney disorders. Beneficial and real-time data readout for preventive diagnosis and clinical evaluation of chronic kidney diseases (CKD) at different stages and renal dysfunction can be acquired by noninvasive monitoring of the creatinine amounts in saliva. The direct determination and real-time response of salivary creatinine can be attained using the supercapacitor-based sensor of cuprous oxide nanoparticles entrapped by the synergistically cross-linked poly(acrylic acid) (PAA) gel-Cu2+ and Nafion perfluorinated membrane fabricated on a screen-printed carbon electrode (SPCE). Here, we demonstrated that the degree of renal illness could be evaluated using salivary creatinine detection via a catalytic mechanism as Cu2+ ions bound irreversibly with C═N functional groups of creatinine. Besides, the computer simulation was performed to study the interaction between 5 functional groups of creatinine toward acrylic gel-Cu2+. The linear increment between the obtained anodic currents and creatinine concentrations varying from 1 to 2000 µM was accomplished with a selectivity efficiency of 97.2%. Nyquist plots obtained by electrochemical impedance spectroscopy (EIS) validated that the increment of impedance changes strongly dependent on the amount of detected creatinine both in artificial and in human saliva. The porosity features were observed in this interconnected nanocomposite and correlated with Nafion doping. Successively, the friendly portable device was invented and integrated saliva sampling with miniaturized, low-cost IoT electronics of world-location mapping, representing the first remote medical sensor focusing on salivary creatinine sensing.

Evidence of Cu(I) Coupling with Creatinine Using Cuprous Nanoparticles Encapsulated with Polyacrylic Acid Gel-Cu(II) in Facilitating the Determination of Advanced Kidney Dysfunctions.

An electrochemical-based sensor created for creatinine detection has been developed for early point-of-care (POC) of diagnosis of renal illnesses. Useful information for the preventive diagnosis and clinical treatments of congenital disorders of creatinine mechanism, advanced liver and kidney diseases, and renal dysfunction can be obtained by the noninvasive evaluation of the creatinine levels in urine. The direct detection of creatinine can be achieved using the modified nanocomposite of cuprous nanoparticles encapsulated by polyacrylic acid (PAA) gel-Cu(II) fabricating on a screen-printed carbon electrode. Here, we report that the degree of kidney dysfunction failure can be determined by an amount of Cu(I) bound with the creatinine through the adsorptive mechanism on the modified electrode. Under cyclic voltammetry scans, the amount of creatinine was measured from the adsorptive signals of the redox peak current identifying the Cu(I)-creatinine complex with a natural logarithm of the creatinine concentration ranging from 200 µM to 100 mM. For this detection range, the theoretical calculation was postulated to describe experimental behaviors of the adsorptive mechanism as creatinine diffused to adsorb on the composite-modified electrode to reduce oxidized copper nanoparticles and transformed to Cu(II)-creatinine complexes. Interestingly, there was evidence that anodic peak potentials had been reduced in magnitudes and shifted negatively by natural logarithm during the formation of the Cu(I)-creatinine complex. For practical usage in POC technology, the creatinine detection in interference was carried out using differential pulse voltammetry to solely determine faradaic currents of creatinine-copper formation. With the interference of urea, glucose, ascorbic acid, glycine, and uric acid in artificial urine, the sensor showed promising results of the interference-free determination with 99.4% sensitivity efficiency, whereas for human urine interference, this sensor showed 85% sensitivity efficiency in detecting creatinine. This shows that this composite-modified sensor (PAA gel-Cu(II)/Cu2O NPs) has great potential for use in the next-generation devices for creatinine sensing to determine the progression in kidney dysfunctions.

Development of a multiplex immunochromatographic strip test and ultrasensitive electrochemical immunosensor for hepatitis B virus screening.

This research proposes two methods for hepatitis B diagnosis including rapid testing and electrochemical assay. For the first method, a multiplex hepatitis B test strip was fabricated to serve as a rapid test for hepatitis B screening. It was developed to simultaneously test three essential serological markers of hepatitis B virus infection including hepatitis B surface antigen (HBsAg), hepatitis B surface antibody (Anti-HBs) and hepatitis B core antibody (Anti-HBc). Gold nanoparticles (GNPs) were used as the signal generator on the test strip. Furthermore, a part of a paper network was incorporated on the strip for the gold-silver enhancement process. This paper network helped in decreasing the analysis time of enhancement and makes the enhancement process easier for rapid testing. The developed test strip was specific for each serological marker. The detection limits of HBsAg, Anti-HBs and Anti-HBc were obtained at 0.5, 0.3 and 0.1 µg mL-1, respectively. For the second method, electrochemical impedance spectroscopy (EIS) was applied for HBsAg detection. This method was proposed for quantitative hepatitis B detection. Anti-HBs antibodies were immobilized on a carbon screen printed electrode (SPCE) via the N-ethyl-N'-(3-(dimethylamino)propyl)carbo-diimide/N-hydroxy succinimide (EDC/NHS) couple reaction which reacted with the carboxyl group of the BSA cross-linked film on the electrode. The electrode modification process was characterized by EIS. A linear relationship between delta charge transfer resistance (ΔRct) and HBsAg concentration was obtained in the range of 5-3000 ng mL-1 with a detection limit of 2.1 ng mL-1. This work is appropriate for quantitative analysis because it is a simple and low-cost method to implement as the SPCE is disposable. Therefore, we hope that this research will be useful to improve hepatitis B detection in the future.

Electrochemical immunoassay platform for high sensitivity protein detection based on redox-modified carbon nanotube labels.

We report a highly sensitive immunoassay protocol based on the use of redox-modified multi-walled carbon nanotubes (MWNTs) as electrochemical labels. The MWNTs were coated with methylene blue (MB) at an optically-determined loading of 3.41 × 10(-3) mol g(-1), and were then attached to secondary antibodies (Ab(2)) by adsorption. As a model analyte mouse IgG was collected by primary antibody (Ab(1))-coated magnetic beads. Following binding of the MB-MWNT-Ab(2) conjugates, IgG could be measured by MB reduction. Using differential pulse voltammetry for quantification, IgG was calibrated with a dynamic range of 0.1 pg mL(-1) to 100 pg mL(-1). Given the different possible Ab(1)-MB-MWNT-Ab(2) orientations on the magnetic beads, it was likely that not all the MB communicated with the electrode. A greater quantity of MB could be accessed by using the Fe(CN)(6)(3-/4-) redox couple as a solution phase mediator. This enabled us to lower the dynamic range down to 5 fg mL(-1) to 100 fg mL(-1).