Reversible detection of proteases and their inhibitors by a pulsed chronopotentiometric polyion-sensitive electrode.
Reversible detection of proteases and their inhibitors by a pulsed chronopotentiometric polyion-sensitive electrode.
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DOI：
10.1016/j.ab.2007.10.043
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发表时间：
2008-03
影响因子：
2.9
通讯作者：
Yida Xu;A. Shvarev;S. Makarychev-Mikhailov;E. Bakker
中科院分区：
文献类型：
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作者：
Yida Xu;A. Shvarev;S. Makarychev-Mikhailov;E. Bakker
脉冲计时电位法操作的聚合物膜电极最近已被引入以取代传统的离子选择性电极用于许多应用。虽然已经报道了用于聚阳离子鱼精蛋白的离子选择性电极，例如，脉冲计时电位读出模式（在此称为脉冲计时电位（pulstrode））提供了改进的稳定性和再现性，同时表现出用于直接检测未稀释的全血样品中的鱼精蛋白的足够的选择性。在这里，这样的鱼精蛋白敏感pulstrodes应用于实时检测的蛋白酶胰蛋白酶及其大豆抑制剂的活性。这是可能的，因为胰蛋白酶消化产生的小片段不能被鱼精蛋白传感膜检测到。实时响应的蛋白水解反应显示出良好的再现性和可逆性，和初始反应速率是依赖于蛋白酶及其抑制剂的浓度。
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.