Toward the Rapid Diagnosis of Sepsis: Detecting Interleukin-6 in Blood Plasma Using Functionalized Screen-Printed Electrodes with a Thermal Detection Methodology

This paper reports the detection of the inflammatory and sepsis-related biomarker, interleukin-6 (IL-6), in human blood plasma using functionalized screen-printed electrodes (SPEs) in conjunction with a thermal detection methodology, termed heat-transfer method (HTM). SPEs are functionalized with antibodies specific for IL-6 through electrodeposition of a diazonium linking group and N'-ethylcarbodiimide hydrochloride (EDC) coupling, which was tracked through the use of cyclic voltammetry and Raman spectroscopy. The functionalized SPEs are mounted inside an additively manufactured flow cell and connected to the HTM device. We demonstrate the ability to detect IL-6 at clinically relevant concentrations in PBS buffer (pH = 7.4) with no significant interference from the similarly sized sepsis-related biomarker procalcitonin (PCT). The limit of detection (3s) of the system is calculated to correspond to 3.4 ± 0.2 pg mL-1 with a working range spanning the physiologically relevant concentration levels in both healthy individuals and patients with sepsis, indicating the sensitivity of the sensor is suitable for the application. Further experiments helped provide a proof-of-application through the detection of IL-6 in blood plasma with no significant interference observed from PCT or the constituents of the medium. Due to the selectivity, sensitivity, straightforward operation, and low cost of production, this sensor platform has the potential for use as a traffic light sensor for the multidetection of inflammatory biomarkers for the diagnosis of sepsis and other conditions in which the rapid testing of blood biomarkers has vital clinical application

Synthesis of PANI/hematite/PB hybrid nanocomposites and fabrication as screen printed paper based sensors for cholesterol detection

In this work, the composites of polyaniline/nano-hematite (α-Fe2O3)/Prussian Blue (PB) were successfully synthesized via a sonochemical method. This nanocomposite (PB/CPANI) has been used for the modification of paper-based sensors for cholesterol detection. The electrochemical studies of this nanocomposite showed a well-defined cyclic voltammogram for hydrogen peroxide (H2O2) with a remarkable electrochemical sensitivity. This nanocomposite modified paper-based electrode also showed excellent electrocatalytic activity towards H2O2 in the interference-free cathodic region. In addition, cholesterol oxidase was immobilized on the PB/CPANI-modified paper-based electrode for selective detection of cholesterol. Under optimum conditions, a linear range of 0.6–6.0 mM with a very low detection limit of 0.52 mM for cholesterol and a good sensitivity of 411.7 μA mM−1 cm−2 were obtained which indicated that the method presented is outstandingly appropriate to determine cholesterol in bovine serum albumin. This work might be an alternative, interference-free, and cost-effective approach for biomonitoring of cholesterol both in methodological studies and in clinical laboratories

On-chip microfluidic systems for determination of L-glutamate based on enzymatic recycling of substrate

Two microfluidic systems have been developed for specific analysis of L-glutamate in food based on substrate recycling fluorescence detection. L-glutamate dehydrogenase and a novel enzyme, D-phenylglycine aminotransferase, were covalently immobilized on (i) the surface of silicon microchips containing 32 porous flow channels of 235 μm depth and 25 μm width and (ii) polystyrene Poros™ beads with a particle size of 20 μm. The immobilized enzymes recycle L-glutamate by oxidation to 2-oxoglutarate followed by the transfer of an amino group from D-4-hydroxyphenylglycine to 2-oxoglutarate. The reaction was accompanied by reduction of nicotinamide adenine dinucleotide (NAD+) to NADH, which was monitored by fluorescence detection (εex=340 nm, εem=460 nm). First, the microchip-based system, L-glutamate was detected within a range of 3.1–50.0 mM. Second, to be automatically determined, sequential injection analysis (SIA) with the bead-based system was investigated. The bead-based system was evaluated by both flow injection analysis and SIA modes, where good reproducibility for L-glutamate calibrations was obtained (relative standard deviation of 3.3% and 6.6%, respectively). In the case of SIA, the beads were introduced and removed from the microchip automatically. The immobilized beads could be stored in a 20% glycerol and 0.5 mM ethylenediaminetetraacetic acid solution maintained at a pH of 7.0 using a phosphate buffer for at least 15 days with 72% of the activity remaining. The bead-based system demonstrated high selectivity, where L-glutamate recoveries were between 91% and 108% in the presence of six other L-amino acids tested