OT-H_2O_2-HRP伏安酶联免疫分析新体系

Investigation of Voltammetric Enzyme-linked Immunoassay Based on New System of OT-H_2O_2-HRP

本文首次提出邻联甲苯胺(OT)-H_2O_2-辣根过氧化物酶(HRP)伏安酶联免疫分析新体系.本方法以线性扫描二阶导数伏安法检测HRP催化H_2O_2氧化OT的产物,用于游离HRP和各种HRP标记物测定,灵敏度比经典的ELISA光度法分别高两个至四个数量级.测定游离HRP的检测限达到1.8×10^(-12)g/mL,线性范围为5.0×10^(-12)～1.0×l0^(-8)g/mL.对此伏安酶联免疫分析新体系的偶合反应机理及电极还原过程也进行了详细的研究.

A voltammetric enzyme - linked immunoassay based on a new system of o- tilidine (OT) - H2O2 - horseradish peroxidase (HRP) has firstly been developed. In this method, the enzyme - catalyzing reaction of H2O2 oxidizing OT couples the electro - reduction reaction of the oxidizing product of OT, which produces a sensitive voltammetric wave at potential of - 0. 58V (vs. SCE) in Britton - Robinson buffer solution. When it is used in the detection of free HRP and labelled HRP, the sensitivity is nearly two and four - order higher than that of classical ELISAmethod,respectively. The detection limit to HRP is 1.8 × 10-12g/mL and the linear range 5.0 ×10-12~ 1.0 × 10-8g/mL. The mechanism of the voltammetric enzyme - linked immunoassay system has also been studied in detail.