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Specifically, we realized rapid binding of bacterial cells to a G-FET by electrical field guiding to the device to realize an overall 3 orders of magnitude decrease in cell-concentration enabling a single-cell detection limit, and 9-fold reduction in needed time to 5 min. Utilizing our new biosensor and procedures, we demonstrate the first selective, electrical detection of the pathogenic bacterial species Staphylococcus aureus and antibiotic resistant Acinetobacter baumannii on a single platform. The modelling of protein-protein binding kinetics is important for the development of affinity-sensors and the prediction of signaling protein based drug efficiency. Therefore, in this research we have evaluated the binding kinetics of several genetically designed protein models (i) three different ligands based on granulocyte colony-stimulating factor GCSF homo-dimeric derivatives linked by differed by linkers of different length and flexibility; (ii) an antibody-like receptor (GCSF-R) based on two GCSF-receptor sites immobilized to Fc domains, which are common parts of protein structures forming antibodies. Genetically engineered GCSF-R is similar to an antibody because it, like the antibody, has two binding sites, which both selectively bind with GCSF ligands. To design the affinity sensor model studied here, GCSF-R was immobilized on a thin gold layer via self-assembled monolayer conjugated with Protein-G. Binding kinetics between immobilized GCSF-R and all three different recombinant GCSF-based homo-dimeric derivatives were evaluated by total internal reflection ellipsometry. Association constants were determined by fitting mathematical models to the experimental data. It was clearly observed that both (i) affinity and (ii) binding kinetics depend on the length and flexibility of the linker that connects both domains of a GCSF-based ligand. The fastest association between immobilized GCSF-R and GCSF-based ligands was observed for ligands whose GCSF domains were interconnected by the longest and the most flexible linker. Here we present ellipsometry-based measurements and models of the interaction kinetics that advance the understanding of bidentate-receptor-based immunosensor action and enables us to predict the optimal linker structure for the design of GCSF-based medications. Thermophoresis is the physical diffusion of molecules from hot to cold induced by a thermal gradient. Thermophoresis has been used to evaluate the interaction of biomolecules in solution. selleck kinase inhibitor In this study, the outer membrane from E. coli was isolated and used to produce OM particles with a diameter of approximately 100 nm. These prepared OM particles were applied in a thermophoretic immunoassay. First, outer membrane (OM) particles with lipopolysaccharides (LPS) and anti-LPS antibodies were used as a model to demonstrate proof of concept and the difference in E. coli thermophoresis was explained by the changes in the molecular surface area (A) and effective charge (σeff). The hydrodynamic size of the molecules was measured as a changing parameter, molecular surface area (A), by dynamic laser scattering (DLS), and the zeta potential was measured as a changing parameter of effective charge (σeff) and then evaluated by the Soret equation. Using the hydrodynamic size and zeta potential values, the interaction between the antigen (OM particle with LPS) and antibody (anti-LPS antibodies) could be monitored and the results were fitted to the thermophoretic immunoassay using the Soret coefficient and equation. Finally, this OM-based immunoassay was applied to the medical diagnosis of systemic lupus erythematosus. Here, OM particles with Ro and La proteins were used to analyze the autoantibodies in patient and control sera. Thermophoretic immunoassay results were also compared to the fitted analysis using hydrodynamic size and zeta potential values and the Soret coefficient and equation. A point-of-care (POC) device to enable de-centralized diagnostics can effectively reduce the time to treatment, especially in case of infectious diseases. However, many of the POC solutions presented so far do not comply with the ASSURED (affordable, sensitive, specific, user-friendly, rapid and robust, equipment free, and deliverable to users) guidelines that are needed to ensure their on-field deployment. Herein, we present the proof of concept of a self-powered platform that operates using the analysed fluid, mimicking a blood sample, for early stage detection of HIV-1 infection. The platform contains a smart interfacing circuit to operate an ultra-sensitive electrolyte-gated field-effect transistor (EGOFET) as a sensor and facilitates an easy and affordable readout mechanism. The sensor transduces the bio-recognition event taking place at the gate electrode functionalized with the antibody against the HIV-1 p24 capsid protein, while it is powered via paper-based biofuel cell (BFC) that extracts the energy from the analysed sample itself. The self-powered platform is demonstrated to achieve detection of HIV-1 p24 antigens in fM range, suitable for early diagnosis. From these developments, a cost-effective digital POC device able to detect the transition from "healthy" to "infected" state at single-molecule precision, with no dependency on external power sources while using minimal components and simpler approach, is foreseen. Quantifying the microRNA (miRNA) level and manipulating them in complex samples, such as serum, is of intense interest because miRNAs are important diagnostic markers. Here, we demonstrate an optical microfiber integrating of untrasensitive detection function and local photothermal therapy potential. A nanointerface consisting of GO supported Cu2-xS nanoplates presented the localized surface plasmon resonance (LSPR) tuned to be consistent with the operation wavelength of the microfiber transducer. It enhanced the surface energy density of evanescent field, on which the miRNA sensing and therapy occurred. With evanescent field enhancement by the plasmonic nanointerface, the sensor exhibits an ultrahigh sensitivity for detecting microRNA at concentrations ranging from 0.1 aM to 10 pM. It is also capable of differentiating one-base mismatches of miRNA at ultralow concentrations (as low as 10 aM) in serum. The photothermal effect of nanointerface simultaneously endows the sensor with the potential for localized photothermal therapy.
Read More: https://www.selleckchem.com/products/ziritaxestat.html
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