使用脂质体免疫裂解测定法（LILA）对血清中C反应蛋白进行均相测定。

Homogeneous determination of C-reactive protein in serum using liposome immune lysis assay (LILA).

作者信息

Umeda M, Ishimori Y, Yoshikawa K, Takada M, Yasuda T

出版信息

Jpn J Exp Med. 1986 Feb;56(1):35-42.

Abstract

Recently, we have developed an analytical system based on a complement dependent liposome immune lysis assay (LILA) to measure antibody against protein antigens. This paper describes the application of LILA to measure protein antigens by inhibition reaction. We choose human C-reactive protein (CRP), one of acute phase reactants, for a model of antigen determination. CRP antigen is coupled to the carboxyfluorescein entrapped multilamellar liposomes by using N-hydroxysuccinimidyl 3-(2-pyridyldithio) propionate and dithiothreitol, and a specific lysis of liposomes is achieved upon the addition of anti-CRP antibody in the presence of complement. Inhibition of this assay can be observed by the addition of competing amounts of CRP in human serum. This inhibition assay is simple, fast, highly sensitive, reproducible and homogeneous. The assay covers the ranges 10-500 micrograms/l. In an experiment using 50 samples, the results by LILA correlated well with those by single radial immunodiffusion and enzyme immunoassay (correlation coefficient: 0.99 and 0.93, respectively).

摘要

最近，我们开发了一种基于补体依赖性脂质体免疫裂解测定法（LILA）的分析系统，用于检测针对蛋白质抗原的抗体。本文描述了LILA通过抑制反应来检测蛋白质抗原的应用。我们选择急性期反应物之一的人C反应蛋白（CRP）作为抗原测定的模型。通过使用N-羟基琥珀酰亚胺3-（2-吡啶二硫基）丙酸酯和二硫苏糖醇，将CRP抗原偶联到包封有羧基荧光素的多层脂质体上，在补体存在下加入抗CRP抗体后可实现脂质体的特异性裂解。在人血清中加入竞争量的CRP可观察到该测定法的抑制作用。这种抑制测定法简单、快速、高度灵敏、可重复且均一。该测定法的范围为10 - 500微克/升。在一项使用50个样本的实验中，LILA的结果与单向放射免疫扩散法和酶免疫测定法的结果相关性良好（相关系数分别为0.99和0.93）。