Solid-cantact electrochemical polyion sensors for monitoring peptidase activities
Solid-cantact electrochemical polyion sensors for monitoring peptidase activities
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DOI：
10.1021/ac701775n
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发表时间：
2008-02-01
影响因子：
7.4
通讯作者：
Shvarev, Alexey
中科院分区：
文献类型：
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作者：
Fordyce, Katherine;Shvarev, Alexey
我们在这里报告改进的固体接触电化学聚离子传感器检测聚离子鱼精蛋白。聚合物膜传感器的制作与导电聚合物作为离子-电子转换层。我们观察。降低所施加的电流脉冲的幅度导致传感器对低鱼精蛋白水平的灵敏度的显著提高。鱼精蛋白传感器表现出稳定和可逆的鱼精蛋白浓度范围从0.05至30毫克L-1的响应。传感器用于监测肽酶的活动，利用恒电流控制的固体接触膜传感器。以聚离子鱼精蛋白为底物，检测蛋白酶胰蛋白酶的活性。通过测量鱼精蛋白浓度来连续监测酶活性，因为鱼精蛋白被酶切割成传感器对其不太敏感的较小片段。当鱼精蛋白浓度在0-5U/mL~（-1）范围内时，反应的起始速率与胰蛋白酶活性呈线性关系。专门检查了对酶反应产物精氨酸的干扰。
We report here on improved solid-contact electrochemical polyion sensors for the detection of polyion protamine. The polymeric membrane sensors were fabricated with a conducting polymer as an ion-electron transduction layer. We observed. that decreasing the magnitude of the applied current pulse caused a significant improvement of the sensor sensitivity to low protamine levels. The protamine sensors exhibited a stable and reversible response to protamine concentrations ranging from 0.05 to 30 mg L-1. The sensors were used for monitoring peptidase activities utilizing galvanostatically controlled solid-contact membrane sensors. The polyion protamine was used as a substrate to detect the activity of the protease trypsin. The enzyme activity was continuously monitored by measuring the protamine concentration as it is cleaved by enzyme into smaller fragments to which the sensor is less sensitive. In the presence of a given level of protamine the initial rate of reaction can be linearly related to the trypsin activity within a 0-5 U mL(-1) concentration range. The interference with the enzymatic reaction product arginine was specifically examined.