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Anaerobic Protein Purification and Kinetic Analysis via Oxygen Electrode for Studying DesB Dioxygenase Activity and Inhibition
Published on: October 3, 2018
An approach to in situ detection of hydrogen peroxide: application of a commercial needle-type electrode
Monika Tamasko1, Livia Nagy, Esztella Mikolas
1Second Department of Medicine and Nephrological Center, Faculty of Medicine, University of Pécs, Pacsirta u. 1, H-7624 Pecs, Hungary.
Abstract:
The reactive oxygen species are thought to play major roles in developing different physiological disorders. A commercial, needle-type amperometric glucose enzyme sensor manufactured for human patients was investigated. This sensor measures glucose by detecting hydrogen peroxide evolved in the enzymatic reaction of glucose. In the experiments, the immobilized enzyme layer of the sensor was inactivated. The applicability of this 'inhibited' glucose sensor for detecting hydrogen peroxide was tested. The simple battery powered, single purpose electronic unit was replaced by an advanced electrochemical workstation. The sensitivity, selectivity and lower limit of detection of the hydrogen peroxide measurements were investigated. Voltammetric measurements were carried out in intensively stirred buffered aqueous media, in plasma samples as well as in subcutan areas of anesthetized Wistar rats. Preliminary measurements carried out with the amperometric and periodically interrupted amperometric technique predicted that the human clinical sensor, after our enzyme inhibition step, can be used for checking the elevation of the hydrogen peroxide level in different subcutan areas of human subjects.
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