甲基红-H_2O_2-HRP伏安酶联免疫分析法测定HRP及其标记物

Methyl red-H_2O_2-HRP voltammetric enzyme-linked immunoassay system for determination of HRP and labels of HRP

提出了一种新的辣根过氧化物酶的底物-甲基红,它本身具有电化学活性,能够在静汞电极上发生还原反应,产生灵敏的伏安电流信号.以H2O2为氧化剂,HRP能催化氧化还原反应的发生,使甲基红被氧化分解,其平衡浓度降低,对应的还原峰电流降低,峰电流的降低值与HRP的质量浓度在5.0×10^-8～5.0×10^-7g/mL之间呈线性关系,对2.0×10^-7g/mL HRP进行11次测定的相对标准偏差为4.6%,方法的检出限为1.8×10^-8g/mL.应用于IgG-HRP和Avidin-HRP的测定.

Vohammetric behaviour of methyl red （MR） in Briton-Robinson buffer solution （ pH 4.35） has been studied in this paper. A new electrochemical system for horseradish peroxidase-MR is established based on the HRP catalyzing redox reaction of MR and H2O2 . The addition of HRP can greatly promote the oxidation reaction. After the addition of HRP, the decrease of the peak current of MR is in proportion to the concentration of HRP. The linear range for the determination of HRP is 5.0 × 10^-8 - 5.0 × 10^-7 g/mL and the detection limit is 1.8 × 10^-8 g/mL. It has been applied to determine the IgG-HRP and avidin-HRP, the highest dilution of avidin-HRP using this system is 1：50000.