摘要
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.
Tn this paper, a new chemiluminescence method for measuring free HRP and labeled HRP is developed by coupling the reaction of H_2O_2-KI catalyzed by HRP with the chemiluminescence reaction of luminol-I_2. The detection limit of HRP is 7 pg/tube and the RSD for 20pg/tube HRP(n=11) is 7.5%. The method hold the linear relationship for HRP over 10-6000 pg/tube.
Comparison of coupling reaction chemiluminescence method and the direct luminol-H_2O_2 chemiluminescence method of HRP shows that the coupling reaction method is 3 times and 10-100 times more sensitive for free HRP and for labeled HRP. The mechanism and kineties characteristic of the coupling reaction has been studied.
出处
《化学学报》
SCIE
CAS
CSCD
北大核心
1991年第4期389-393,共5页
Acta Chimica Sinica
基金
国家自然科学基金
