以3,3′,5,5′-四甲基联苯胺为底物的伏安酶联免疫分析体系研究

Voltammetric Enzyme-linked Immunoassay System Using 3,3′,5,5′-Tetramethyl Benzidine as Substrate

对 3 ,3′,5 ,5′ 四甲基联苯胺 (TMB) H2 O2 辣根过氧化物酶 (HRP)伏安酶联免疫分析体系进行了研究。HRP催化H2 O2 氧化TMB ,其氧化产物在汞电极上可以发生还原反应 ,在B R缓冲溶液中 ,-0 .76V(vs.SCE)处产生较为灵敏的伏安波。将此伏安波应用于测定HRP和HRP的标记物。测定HRP的线性范围为 6.0×1 0 - 1 1 ～ 5 .0× 1 0 - 9g mL ,检测限为 5 .0× 1 0 - 1 1 g mL。

A voltammetric enzyme-linked immunoassay system using 3,3', 5, 5-tetramethyl benzidine (TMB) as substrate has been investigated. With this method, the horseradish peroxidase (HRP)-catalyzed reaction of H2O2 oxidizing TMB is coupled with the electro-reduction reaction of the oxidized product of TMB on the mercury electrode, which produces a sensitive voltammetric reductive wave at the potential of -0.76 V (vs. SCE) in Britton-Robinson (B-R) buffer solution. When it was used in the detection of free HRP, the detection limit of HRP was 5.0 x 10(-11) g/mL with the linear range from 6.0 x 10(-11) to 5.0 x 10(-9) g/mL. The mechanism of this voltammetric enzyme immunoassay system has also been studied.