使用戊二醛不溶性抗体制剂重新分离免疫反应性放射性碘化抗原。

Reisolation of immunoreactive radioiodinated antigens using glutaraldehyde-insolubilized antibody preparations.

作者信息

Anderson B, Donakowski C, Entwistle R, Davis L

出版信息

J Immunol Methods. 1980;36(3-4):309-14. doi: 10.1016/0022-1759(80)90135-0.

Abstract

Radioiodination of antigens for use in radioimmunoassay can result in substantial losses of antigenic reactivity with the corresponding antibody and antisera preparations. We describe a method whereby antigens iodinated with the chloramine-T procedure are bound to and eluted from glutaraldehyde-insolubilized antibody. Unfractionated antisera, an ammonium sulfate precipitated fraction or the IgG fraction of antisera may be used as insolubilized immunoadsorbents. The method has been applied for the reisolation of a radioiodinated peptide, a low molecular weight protein and the fibronectin molecule. The total binding of 125I-antigens to antibody in radioimmunoassays can be increased from such low amounts before reisolation that the assay is not feasible, to 85% above background binding, after adsorption and elution from the insolubilized antibody preparations.

摘要

用于放射免疫分析的抗原放射性碘化可能会导致与相应抗体及抗血清制剂的抗原反应性大幅丧失。我们描述了一种方法，通过该方法，用氯胺 - T 法碘化的抗原可与戊二醛固定化抗体结合并洗脱。未分级的抗血清、硫酸铵沉淀级分或抗血清的 IgG 级分均可用作固定化免疫吸附剂。该方法已应用于放射性碘化肽、低分子量蛋白质和纤连蛋白分子的再分离。在放射免疫分析中，¹²⁵I - 抗原与抗体的总结合量可从再分离前低到无法进行分析的水平，提高到从固定化抗体制剂吸附和洗脱后高于背景结合 85% 的水平。