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化学学报
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化学学报 ›› 1991, Vol. 49 ›› Issue (4): 389-393. 上一篇    下一篇

研究论文

偶合反应化学发光酶免疫分析研究: I. HRP及其标记物的化学发 光测定

章竹君;拜明岐;张新荣   

  1. 陕西师范大学化学系.西安(710062)
  • 发布日期:1991-04-15

Chemiluminescence enzyme immunoassay with coupling reaction: I. Determination of HRP and labeled HRP by chemiluminescence analysis

Zhang Zhujun;Bai Mingqi;Zhang Xinrong   

  1. Shanxi Normal Univ, Dept Chem.Xian(710062)
  • Published:1991-04-15

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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Luminol-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。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。

关键词: 测定, 化学发光, 酶免疫分析, 过氧化氢, 碘化钾, 辣根过氧化物酶, 抗体, 偶合反应, 机理, 动力学

In 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 limits 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 10-6000pg/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.

Key words: DETERMINATION, CHEMILUMINESSENCE, HYDROGEN PEROXIDE, POTASSIUM IODIDE, ANTIBODY, COUPLING REACTION, MECHANISM, DYNAMICS

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