竞争性酶联免疫吸附测定（ELISA）的简单理论处理及其在人类血型抗原检测中的应用。

A simple theoretical treatment of a competitive enzyme-linked immunosorbent assay (ELISA) and its application to the detection of human blood group antigens.

作者信息

Chapman V, Fletcher S M, Jones M N

机构信息

Department of Biochemistry and Molecular Biology, School of Biological Sciences, University of Manchester, U.K.

出版信息

J Immunol Methods. 1990 Jul 20;131(1):91-8. doi: 10.1016/0022-1759(90)90237-p.

DOI:10.1016/0022-1759(90)90237-p

PMID:2380571

Abstract

A theory has been developed to explain the behaviour of a competitive enzyme-linked immunosorbent assay (ELISA) in which an immobilized antigen competes with a liquid-phase antigen for a limiting amount of antibody. The binding of antibody to antigen on a solid surface (microtitre well) is described in terms of Langmuirian adsorption with a binding constant kappa. Two equations are presented to describe the behaviour of the ELISA signal as a function of competing antigen concentration; an exact equation and an approximate equation which can be used when the surface coverage of the immobilized antigen is not known. It is shown how curves of ELISA signal vs. competing antigen concentration depend on K/kappa [antibody]. The theory has been tested using several immobilized blood group A antigens competing with ovarian cyst fluid A substance and found to adequately describe these competitive ELISAs which have a detection limit of approximately 1 ng of blood group antigen.

摘要

已经建立了一种理论来解释竞争性酶联免疫吸附测定（ELISA）的行为，在这种测定中，固定化抗原与液相抗原竞争有限量的抗体。根据具有结合常数κ的朗缪尔吸附来描述抗体与固体表面（微量滴定孔）上抗原的结合。给出了两个方程来描述ELISA信号随竞争抗原浓度的变化行为；一个精确方程和一个近似方程，当固定化抗原的表面覆盖率未知时可以使用。展示了ELISA信号与竞争抗原浓度的曲线如何取决于K/κ[抗体]。该理论已通过使用几种固定化的A血型抗原与卵巢囊肿液A物质竞争进行了测试，发现能够充分描述这些竞争性ELISA，其检测限约为1 ng血型抗原。