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Amperometric immunosensor for diagnosis of albuminuria

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MIKUŠOVÁ Zuzana PASTUCHA Matěj FARKA Zdeněk ČUNDERLOVÁ Veronika SKLÁDAL Petr

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Fakulta / Pracoviště MU

Středoevropský technologický institut

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Popis Human serum albumin (HSA) is a plasma protein involved in transportation of ligands to specific targets. Its increased presence in urine (microalbuminuria) is considered as an indication of diabetic nephropathy. Currently, HSA is being typically detected by immunoassays such as ELISA. These methods provide high sensitivity; however, they are time consuming, complicated and expensive. One of the possibilities to over-come these disadvantages is to use electrochemical biosensors that offer operational simplicity, short analysis times and miniaturization possibilities for point-of-care diagnosis. The electrochemical biosensors are typically based on screen-printed electrodes (SPEs) due to their low cost, portability and mass production capabilities. We have developed an amperometric immunosensor for rapid diagnostics of albumi-nuria. Anti-HSA antibody was immobilized to the surface of SPEs that allowed specific capture of HSA. The detection was based on binding of secondary antibody conjugated with horseradish peroxidase (HRP), that provides oxidation of 3,3´,5,5´-tetramethylbenzidine (TMB) in presence of H2O2. Alternatively, Prussian blue catalytic nanoparticles were employed as convenient enzyme replacement. SPEs based on gold and carbon (including carbon nanotubes) working electrode were compared. This method has proved to be suitable for determination of clinically relevant concentrations of HSA in urine, optimization focused on better detection limit is in progress.
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