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本文将辣根过氧化物酶(HRP)催化过氧化氢氧化KI生成I_2的反应与Lumnol-I_2的化学发光反应相偶合,提出了测定HRP及其抗原、抗体标记物的高灵敏度的化学发光法.检测下限为7pg,线性范围为10—6000pg.较通常使用的HRP直接催化Luminol-H_2O_2化学发光法灵敏度高10—100倍;克服了固相吸附法直接测定HRP标记物的缺陷,提高了测定的选择性.文中还对该偶合反应的机理及其动力学特性进行了研究.
In this paper, the reaction of horseradish peroxidase (HRP) catalyzed oxidation of KI by H2O2 to I_2 was coupled with the chemiluminescence reaction of Lumnol-I_2, and the chemiluminescence method for the determination of HRP and its antigens and antibody markers was proposed .The detection limit was 7 pg and the linear range was 10-6000 pg.The sensitivity of Luminol-H_2O_2 chemiluminescence method was 10-100 times higher than that of the commonly used HRP direct catalysis.It overcomes the defects of direct determination of HRP markers by solid-phase adsorption and improved detection The mechanism of this coupling reaction and its kinetic properties are also studied.