Reversible detection of proteases and their inhibitors by a pulsed chronopotentiometric polyion-sensitive electrode
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Xu, Yida and Shvarev, Alexey and Makarychev-Mikhailov, Sergey and Bakker, Eric (2008) Reversible detection of proteases and their inhibitors by a pulsed chronopotentiometric polyion-sensitive electrode, Analytical Biochemistry 374(2):366-370.
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Polymer membrane electrodes operated by pulsed chronopotentiometry have recently been introduced to replace traditional ion-selective electrodes for a number of applications. While ion-selective electrodes for the polycation protamine have been reported, for instance, a pulsed chronopotentiometric readout mode (called here pulstrode) provides improved stability and reproducibility while exhibiting sufficient selectivity for the direct detection of protamine in undiluted whole blood samples. Here, such protamine-sensitive pulstrodes are applied for the real-time detection of the activity of the protease trypsin and its soybean inhibitor. This is possible because small fragments produced by the trypsin digestion are not detectable by the protamine-sensing membrane. The real-time response to the proteolytic reaction is shown to exhibit good reproducibility and reversibility, and the initial reaction rate is dependent on the concentration of the protease and its inhibitor.
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