Sumner C, Krause S, Sabot A, Turner K, McNeil C J
Department of Chemistry, University of Sheffield, Sheffield S3 7HF, UK.
Biosens Bioelectron. 2001 Dec;16(9-12):709-14. doi: 10.1016/s0956-5663(01)00210-x.
A biosensor based on the enzyme-catalysed dissolution of biodegradable polymer films has been developed. Three polymer-enzyme systems were investigated for use in the sensor: a poly(ester amide), which is degraded by the proteolytic enzyme alpha-chymotrypsin; a dextran hydrogel, which is degraded by dextranase; and poly(trimethylene) succinate, which is degraded by a lipase. Dissolution of the polymer films was monitored by Surface Plasmon Resonance (SPR). The rate of degradation was directly related to enzyme concentration for each polymer/enzyme couple. The poly(ester amide)/alpha-chymotrypsin couple proved to be the most sensitive over a concentration range from 4 x 10(-11) to 4 x 10(-7) mol l(-1) of enzyme. The rate of degradation was shown to be independent of the thickness of the poly(ester amide) films. The dextran hydrogel/dextranase couple was less sensitive than the poly(ester amide)/alpha-chymotrypsin couple but showed greater degradation rates at low enzyme concentrations. Enzyme concentrations as low as 2 x 10(-11) mol l(-1) were detected in less than 20 min. Potential fields of application of such a sensor system are the detection of enzyme concentrations and the construction of disposable enzyme based immunosensors, which employ the polymer-degrading enzyme as an enzyme label.
一种基于酶催化可生物降解聚合物薄膜溶解的生物传感器已被开发出来。研究了三种聚合物 - 酶系统用于该传感器:一种聚(酯酰胺),可被蛋白水解酶α - 胰凝乳蛋白酶降解;一种葡聚糖水凝胶,可被葡聚糖酶降解;以及聚(三亚甲基)琥珀酸酯,可被脂肪酶降解。通过表面等离子体共振(SPR)监测聚合物薄膜的溶解情况。对于每对聚合物/酶,降解速率与酶浓度直接相关。在酶浓度范围为4×10^(-11)至4×10^(-7) mol l^(-1)时,聚(酯酰胺)/α - 胰凝乳蛋白酶对被证明是最敏感的。降解速率显示与聚(酯酰胺)薄膜的厚度无关。葡聚糖水凝胶/葡聚糖酶对比聚(酯酰胺)/α - 胰凝乳蛋白酶对敏感性较低,但在低酶浓度下显示出更高的降解速率。在不到20分钟内就能检测到低至2×10^(-11) mol l^(-1)的酶浓度。这种传感器系统的潜在应用领域包括酶浓度的检测以及基于聚合物降解酶作为酶标记的一次性酶基免疫传感器的构建。