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[过氧化物酶及其缀合物与溶液中特异性抗体相互作用的平衡和动力学参数]

[Equilibrium and kinetic parameters of the interaction of peroxidase and its conjugates with specific antibodies in solution].

作者信息

Girina N V, Karaseva E I, Metelitsa D I

出版信息

Biokhimiia. 1994 Dec;59(12):1864-71.

PMID:7873686
Abstract

Horseradish peroxidase (HRP) and its conjugates with IgG (HRP-IgG) and cortisol (HRP-COR) interactions with polyvalent antibodies against peroxidase (ANTI-HRP) have been studied by homogeneous enzyme immunoassay. The total catalytic activity in o-phenylenediamine oxidation for free peroxidase or its conjugates and their immunocomplexes with antibodies against the enzyme (ANTI-HRP) has been measured. Linearization of time dependencies of the initial rate for peroxidation in the ln (v-v infinity)-time coordinates, where v and v infinity are the reaction rates at a definite moment of time and after the onset of the equilibrium, as well as in the [ANTI-HRP]/(v-v infinity)-[ANTI-HRP]0 and I/[ANTI-HRP]0k1 + k-1)-[ANTI-HRP]0 made it possible to determine the dissociation constant, Kd, and rate constant for the formation and dissociation, for the immunocomplexes k1 and k-1. The Kd values were in good agreement with those determined by the classical Scatchard method.

摘要

已通过均相酶免疫测定法研究了辣根过氧化物酶(HRP)及其与IgG(HRP-IgG)和皮质醇(HRP-COR)的缀合物与抗过氧化物酶多价抗体(ANTI-HRP)的相互作用。已测量了游离过氧化物酶或其缀合物及其与抗酶抗体(ANTI-HRP)的免疫复合物在邻苯二胺氧化中的总催化活性。在ln(v-v无穷大)-时间坐标中,过氧化物初始速率的时间依赖性线性化,其中v和v无穷大是在特定时刻和平衡开始后的反应速率,以及在[ANTI-HRP]/(v-v无穷大)-[ANTI-HRP]0和I/[ANTI-HRP]0k1 + k-1)-[ANTI-HRP]0中,使得能够确定免疫复合物的解离常数Kd以及形成和解离的速率常数k1和k-1。Kd值与通过经典Scatchard方法测定的值高度一致。

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