Conservation of water resources calls for ever stricter regulatory measures and better monitoring systems. Whole-cell bacterial sensors have been genetically engineered to react to target toxicants by the induction of a selected promoter and the subsequent production of bioluminescent light through a recombinant lux reporter. In order to create a one-step assay, we have designed a new, self-contained, disposable optical fiber sensor module and a customized photodetector system that integrates these microorganisms. A photon-counting photomultiplier tube-based instrument was constructed. Optical fiber tip cores were covered with adlayer films consisting of calcium alginate containing bioluminescent bacterial sensors of genotoxicants. Multiplying these steps thickened the adlayer in increments, increasing the number of bacterial reporters attached to the optical fiber transducer. These whole cell optrodes are responsive to external traces of DNA damaging agents such as mitomycin C. Light production was shown to be dose-dependent and proportional to the number of bacterial layers.
Surface-plasmon-resonance (SPR) is a quantum-electromagnetic phenomenon arising from the interaction of light with free electrons at a metal-dielectric interface. At a specific angle/wavelength of light, the photon’s energy is transferred to excite the oscillation of the free electrons on the surface. A change in the refractive-index (RI) may occur, which is influenced by the analyte concentration in the medium in close contact with the metal surface. SPR has been widely used for the detection of gaseous, liquid, or solid samples. In this study, a functionalized specific SPR chip was designed and used in a novel point-of-care SPR module (PhotonicSys SPR H5) for the detection of the stroke biomarkers NT-proBNP and S100β. These biomarkers have proven to be good for stroke diagnosis, with sensitivity and specificity of >85%. Specific detection was done by binding a biomolecular-recognizing antibody onto the Au SPR-chip. Detection was tested in water and plasma samples. NT-proBNP and S100β were detected in a range of concentrations for stroke, from 0.1 ng/mL to 10 ng/mL. The RI of the blank plasma samples was 1.362412, and the lowest concentration tested for both biomarkers showed a prominent shift in the RI signal (0.25 ng/mL NT-proBNP (1.364215) and S100β (1.364024)). The sensor demonstrated a clinically relevant limit-of-detection of less than ng/mL.
Paper-based colorimetric biosensors combine the use of paper with colorimetric signal detection. However, they usually demonstrate lower sensitivities because a signal amplification procedure has not been used. Stopping the reaction of colorimetric signal generation is often used in lab-based assays in order to amplify and stabilize the colorimetric signal for detection. In this study, the generation of a stopped colorimetric signal was examined for accurate and enhanced signal detection in paper-based biosensors. The colorimetric reaction in biosensors is usually based on the interaction between the enzyme horseradish peroxidase (HRP) and a selected chromogenic substrate. The two most commonly used HRP substrates, 3,3’,5,5’-tetramethylbenzidine (TMB) and 2’-azinobis (3-ethylbenzothiazoline-6-sulfonic-acid) (ABTS), were compared in terms of their ability to generate a stopped colorimetric signal on membrane. The stopped colorimetric signal was visible for TMB but not for ABTS. Moreover, the generation of stopped colorimetric signal was dependent on the presence of polyvinylidene-difluoride (PVDF) membrane as the separation layer. With PVDF the colorimetric signal (color intensity) was higher (TMB: 126 ± 6 and ABTS: 121 ± 9) in comparison to without PVDF (TMB: 110 ± 2 and ABTS: 102 ± 4). The TMB stopped colorimetric signal demonstrated a more stable signal detection with lower standard deviation values. To conclude, a stopped colorimetric signal can be generated in paper-based biosensors for enhanced and accurate signal detection.
Enzyme mimetics have attracted wide interest due to their inherent enzyme-like activity and unique physicochemical properties, as well as promising applications in disease diagnosis, treatment and monitoring. Inspired by the attributes of nonheme iron enzymes, synthetic models were designed to mimic their capability and investigate the catalytic mechanisms. Herein, metal-organic gels (Fe-MOGs) with horseradish peroxidase (HRP) like Fe-NX structure were successfully synthesized though the coordination between iron and 1,10-phenanthroline-2,9-dicarboxylic acid (PDA) and exhibited excellent peroxidase-like activity. Its structure-activity relationship and the in-situ electrochemiluminescence (ECL) detection of H2O2 secreted by Hela cells were further investigated. The highly dispersed Fe-NX active sites inside Fe-MOGs were able to catalyze the decomposition of H2O2 into large amounts of reactive oxygen species (ROS) via a Fenton-like reaction under a low overpotential. Due to the accumulation of ROS free radicals, the luminol ECL emission was significantly amplified. A proof-ofconcept biosensor was constructed with a detection limit as low as 2.2 nM and a wide linear range from 0.01 to 40 μM. As a novel metal organic gels based enzyme mimetic, Fe-MOGs show great promises in early cancer detection and pathological process monitoring.
Rational design and facile synthesis of highly active electrocatalysts with low cost for oxygen reduction reaction (ORR) are always of great challenge. Specifically, development of a new type of energy-saving materials with convenient method is regarded as the current bottleneck. Herein, an innovative strategy based on amino trimethylene phosphonic acid (ATMP) as chelating agent for cobalt-metaphosphate coordination polymer is reported to one-pot synthesis of a novel precursor in methanol for ORR electrocatalyst. Carbonization of the precursor at 900 o C at N 2 atmosphere results in the feasible formation of cobalt metaphosphate based composite (Co(PO 3) 2 /NC). A further step in the thermal cleavage at 650 o C at air for 4h, Co(PO 3) 2 /NC can be finally transformed into inorganic Co(PO 3) 2. Advanced spectroscopic techniques and density function theory (DFT) calculations are applied to confirm the main catalytically active center and the physical properties of Co(PO 3) 2 /NC. This obtained Co(PO 3) 2 /NC nanocomposite exhibits superior electrocatalysis to Co(PO 3) 2 with an enhanced onset potential (0.906 V vs. RHE) and diffusion limiting current (5.062 mA cm-2), which are roughly close to those of commercial 20 % Pt/C (0.916 V, 5.200 mA cm-2).
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