Fernando S A, Wilson G S
Department of Chemistry, University of Kansas, Lawrence 66045.
J Immunol Methods. 1992 Jul 6;151(1-2):47-66. doi: 10.1016/0022-1759(92)90104-2.
The one-step sandwich immunoassay is increasingly replacing the traditional two-step immunoassay due to obvious advantages such as assay speed. However, the one-step sandwich immunoassay suffers from the 'hook' effect irrespective of the analyte characteristics. The 'hook' effect is dependent primarily on the analyte concentration. Three different model analytes, human growth hormone (hGH), the dimeric form of hGH (D-hGH, having a discrete number of repeating epitopes) and ferritin (multiple epitopes) having different immunological properties have been employed in studies of the one-step sandwich immunoassay. The characteristics of each of the model analytes offer new insights into general guidelines for assay procedures. These guidelines permit rapid optimization of assay conditions for an immunoassay without a priori knowledge of the immunological characteristics of the antibody or antigen. Both experimental and theoretical data show several instances where high capacity solid-phase antibodies can effectively shift the 'hook' to relatively higher analyte concentrations. The effect of the concentration of labeled antibody on assay response was examined theoretically.
一步夹心免疫测定法因其在检测速度等方面的明显优势，正日益取代传统的两步免疫测定法。然而，无论分析物的特性如何，一步夹心免疫测定法都会受到“钩状效应”的影响。“钩状效应”主要取决于分析物的浓度。在一步夹心免疫测定法的研究中，使用了三种具有不同免疫特性的不同模型分析物，即人生长激素（hGH）、hGH的二聚体形式（D-hGH，具有离散数量的重复表位）和铁蛋白（多个表位）。每种模型分析物的特性为检测程序的一般指南提供了新的见解。这些指南允许在无需事先了解抗体或抗原免疫特性的情况下，快速优化免疫测定的检测条件。实验和理论数据均表明，在多个实例中，高容量固相抗体可有效将“钩状效应”转移至相对较高的分析物浓度。从理论上研究了标记抗体浓度对检测响应的影响。
