Ortega-Vinuesa J L, Molina-Bolívar J A, Peula J M, Hidalgo-Alvarez R
Departamento de Física Aplicada, Facultad de Ciencias, Universidad de Granada, Spain.
J Immunol Methods. 1997 Jul 14;205(2):151-6. doi: 10.1016/s0022-1759(97)00064-1.
This work is based on the well-established immunoassay principle of agglutination of latex particles covered by immunoproteins. In our experiments, positively charged particles act as carriers for the F(ab')2 fragment, obtained from rabbit polyclonal IgG, active against C-reactive protein (CRP). The presence of the antigen CRP in the immunolatex system causes agglutination and the aim of the present study was to compare different optical techniques (turbidimetry, nephelometry, angular anisotropy and photon correlation spectroscopy) capable of detecting the agglutination. The sensitivity and detection limit largely depend on the optical method. We have analyzed for each optical technique the following aspects: sensitivity, reproducibility, detection limit, reaction time, amount of sample wasted and availability of the required detection device. The results presented in this paper show that both angular anisotropy and photon correlation spectroscopy offer lower detection limits, and use little reagent, but have longer assay times than the classical optical techniques of turbidimetry and nephelometry.
这项工作基于免疫蛋白覆盖的乳胶颗粒凝集这一成熟的免疫分析原理。在我们的实验中,带正电的颗粒作为F(ab')2片段的载体,该片段从兔多克隆IgG获得,对C反应蛋白(CRP)具有活性。免疫乳胶系统中抗原CRP的存在会导致凝集,本研究的目的是比较能够检测凝集的不同光学技术(比浊法、散射比浊法、角各向异性和光子相关光谱法)。灵敏度和检测限在很大程度上取决于光学方法。我们针对每种光学技术分析了以下方面:灵敏度、重现性、检测限、反应时间、浪费的样品量以及所需检测设备的可用性。本文给出的结果表明,角各向异性和光子相关光谱法都具有较低的检测限,且试剂用量少,但与比浊法和散射比浊法等经典光学技术相比,测定时间更长。