Achtnich U R, Tiefenauer L X, Andres R Y
Paul Scherrer Institute, Project Medical Bioanalytics, Villigen, Switzerland.
Biosens Bioelectron. 1992;7(4):279-90. doi: 10.1016/0956-5663(92)87006-b.
One of the most crucial steps for the successful construction of a biosensor is the appropriate and reproducible coupling of the biological part (e.g. enzyme, antibody) to the inorganic moiety of the device (e.g. electrode, microchip). In this paper three methods of immobilization of avidin to a glassy carbon electrode are described. Depending on the type of immobilization, avidin may lose its biological activity as determined by an enzyme immunoassay, using biotinylated reagents. If avidin is covalently bound to the glassy carbon electrode via the bridge molecule 4.4'-diaminodiphenylamine, the biological activity is retained. About 1.5 pmol of avidin can be bound to the electrode (3 mm in diameter), resulting in a nearly complete monolayer of protein.
生物传感器成功构建的最关键步骤之一是将生物部分（如酶、抗体）与设备的无机部分（如电极、微芯片）进行适当且可重复的偶联。本文描述了三种将抗生物素蛋白固定到玻碳电极上的方法。根据固定类型的不同，通过使用生物素化试剂进行酶免疫测定可知，抗生物素蛋白可能会失去其生物活性。如果抗生物素蛋白通过桥连分子4,4'-二氨基二苯胺共价结合到玻碳电极上，则其生物活性得以保留。约1.5皮摩尔的抗生物素蛋白可结合到电极（直径3毫米）上，形成几乎完整的蛋白质单层。
