Rubio Retama J, Sánchez-Paniagua López M, Hervás Pérez J P, Frutos Cabanillas G, López-Cabarcos E, López-Ruiz B
Departamento de Físico-Química Farmacéutica, Facultad de Farmacia, Universidad Complutense de Madrid, 28040 Madrid, Spain.
Biosens Bioelectron. 2005 May 15;20(11):2268-75. doi: 10.1016/j.bios.2004.10.011.
Acrylic microgels are proposed as enzyme immobilizing support in amperometric biosensors. Two enzymes, glucose oxidase and tyrosinase, were entrapped in this matrix and their behaviour is compared. The optimum cross-linking of the polymeric matrix required to retain the enzyme, and to allow the diffusion of the substrate is different for each enzyme, 3.2% for glucose oxidase and 4.5% for tyrosinase. The effect of pH and temperature on the biosensor responses has been studied by experimental design methodology and predictions have been compared with independently performed experimental measurements. A quadratic effect of the variables studied (pH and T) on the biosensor response and the small or null interaction between them was confirmed. The pH results obtained with both methods are coincident revealing an reversible effect on the enzyme. However, the temperature optimum value obtained by experimental design was 10 degrees C lower as a result of an activity decay due to irreversible thermal denaturation of both enzymes.
丙烯酸微凝胶被提议用作电流型生物传感器中酶固定化的载体。两种酶,葡萄糖氧化酶和酪氨酸酶,被包埋在这种基质中,并对它们的行为进行了比较。保留酶并允许底物扩散所需的聚合物基质的最佳交联度因每种酶而异,葡萄糖氧化酶为3.2%,酪氨酸酶为4.5%。通过实验设计方法研究了pH值和温度对生物传感器响应的影响,并将预测结果与独立进行的实验测量结果进行了比较。研究变量(pH值和温度)对生物传感器响应的二次效应以及它们之间的小或零相互作用得到了证实。两种方法获得的pH值结果一致,表明对酶有可逆影响。然而,由于两种酶不可逆热变性导致活性衰减,通过实验设计获得的最佳温度值低了10摄氏度。