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抗体标记胶体金颗粒的表征及其在溶胶颗粒免疫分析(SPIA)中的应用。

Characterization of antibody labelled colloidal gold particles and their applicability in a sol particle immuno assay (SPIA).

作者信息

Martin J M, Pâques M, van der Velden-de Groot T A, Beuvery E C

机构信息

Laboratory for Inactivated Viral Vaccines, National Institute for Public Health and Environmental Protection, Bilthoven, The Netherlands.

出版信息

J Immunoassay. 1990;11(1):31-47. doi: 10.1080/01971529008053256.

Abstract

This study describes the characterization of antibody labelled colloidal gold particles and their applicability in a sol particle immuno assay (SPIA) to quantify murine monoclonal antibodies of all IgG isotypes. Two physical methods (transmission electron microscopy (TEM) and dynamic light scattering (DLS], were used to obtain information about particle size and morphology of the gold sols, but only with DLS antibody labelling could be detected. In addition, electrophoretic methods like agarose electrophoresis and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) showed also that antibody labelling was successful. The biological activity of the antibody gold conjugates was determined by using them in a SPIA and as an electrondense marker in an immunogold labelling procedure to visualize meningococcal surface exposed outer membrane proteins labelled with monoclonal antibodies. The SPIA was applicable to determine murine monoclonal antibodies of all IgG isotypes with a sensitivity of 20-80 ng/ml and a coëfficiënt of variation of 6.7 +/- 2.2%.

摘要

本研究描述了抗体标记的胶体金颗粒的特性及其在溶胶颗粒免疫分析(SPIA)中用于定量所有IgG同种型小鼠单克隆抗体的适用性。使用两种物理方法(透射电子显微镜(TEM)和动态光散射(DLS))来获取有关金溶胶颗粒大小和形态的信息,但只有通过DLS才能检测到抗体标记。此外,琼脂糖电泳和十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)等电泳方法也表明抗体标记是成功的。通过在SPIA中使用抗体金缀合物,并将其作为免疫金标记程序中的电子致密标记物,以可视化用单克隆抗体标记的脑膜炎球菌表面暴露的外膜蛋白,来确定抗体金缀合物的生物活性。SPIA适用于测定所有IgG同种型的小鼠单克隆抗体,灵敏度为20-80 ng/ml,变异系数为6.7±2.2%。

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