联苯胺-H_2O_2-HRP偶合反应伏安酶联免疫分析新体系测定HRP的研究

Determination of HRP with Coupling Reaction Voltammetric EnzymeLinked Immunoassay Besed on BenzidineH_2O_2HRP System

提出了联苯胺－Ｈ２Ｏ２－ＨＲＰ偶合反应伏安酶联免疫分析新体系测定ＨＲＰ的方法。通过测定ＨＲＰ催化Ｈ２Ｏ２氧化联苯胺的产物间接测定ＨＲＰ的浓度。偶合反应产物在ｐＨ５．２的ＢＲ支持电解质中，在－０．６２Ｖ（ＳＣＥ）附近产生一灵敏的极谱波，在选定的最佳条件下，测定ＨＲＰ的线性范围是５．０×１０－８～１．０×１０－４ｇ／Ｌ，检出限可达７．２×１０－９ｇ／Ｌ。用于测定游离的ＨＲＰ和ＨＲＰ的标记物，灵敏度都比经典的ＥＬＩＳＡ法高。

HRP and labelled HRP have been determined with coupling reaction voltammetric enzymelinked immunoassay besed on benzidineH_2O_2HRP system.HRP catalyses the oxidation of benzidine by H_2O_2,the product of which has a sensitive voltammetric wave at potential of -0.62V (SCE) in pH4.0 HAcNaAc buffer solution.Trace amount of HRP may be determined with a detection limit of 7.2×10-9g/L and a linear range of 5.0×10-8～10×10-4g/L .Sensitivity of this method to determine free HRP and labelled HRP is 4 times higher than that of ELISA method.