Perez-Amodio S, Holownia P, Davey C L, Price C P
St. Bartholomew's and Royal London Hospital's School of Medicine and Dentistry, Department of Clinical Biochemistry, UK.
Anal Chem. 2001 Jul 15;73(14):3417-25. doi: 10.1021/ac001529h.
The role of the solution environment for a light-scattering, latex-particle-enhanced, homogeneous immunoassay of C-reactive protein (CRP) has been investigated in order to assess and optimize the immunoagglutination response. Latex particles of 50-170-nm sizes were covalently coupled with an IgG polyclonal antibody and subjected to an extensive optimization regime. This consisted of conditions responsible, in different degrees, for the principal attractive/repulsive forces affecting both colloidal stability and the antibody/antigen interaction: particle size, antibody concentration, ionic strength and species, pH, and amino acid chemistry of the particle surface. Careful control of these parameters was found to be necessary to achieve the desired effects of balancing high colloidal stability in the absence of antigen but promoting a rapid, sensitive, and dose-dependent agglutination with pathological serum samples. In addition, the estimation of fundamental properties governing intermolecular interaction (i.e. the "Hamaker" constant and critical coagulation concentration) was attempted to order to investigate a simple, practical means of defining a colloidal/immunoassay system under "real conditions" as well as "real time". It is concluded that because each antibody system is unique, a similar optimization should be performed in diagnostic immunoassays of this type to maximize their clinical utility.
为了评估和优化免疫凝集反应，研究了溶液环境在以光散射、乳胶颗粒增强的C反应蛋白（CRP）均相免疫测定中的作用。将50-170纳米大小的乳胶颗粒与IgG多克隆抗体共价偶联，并进行广泛的优化方案。该方案包括在不同程度上影响胶体稳定性和抗体/抗原相互作用的主要吸引/排斥力的相关条件：颗粒大小、抗体浓度、离子强度和种类、pH值以及颗粒表面的氨基酸化学性质。发现仔细控制这些参数对于实现所需效果是必要的，即在没有抗原的情况下实现高胶体稳定性，同时促进与病理血清样本的快速、灵敏和剂量依赖性凝集。此外，尝试估算控制分子间相互作用的基本性质（即“哈梅克”常数和临界凝聚浓度），以便研究在“实际条件”以及“实时”下定义胶体/免疫测定系统的简单实用方法。得出的结论是，由于每个抗体系统都是独特的，因此在这类诊断免疫测定中应进行类似的优化，以最大限度地提高其临床效用。
