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一种基于蛋白A结合、聚乙二醇沉淀的免疫放射分析方法。应用于检测人血清中的免疫复合物和C3以及交叉反应寄生虫提取物中的自身抗原。

A protein A-binding, polyethylene glycol precipitation-based immunoradiometric assay. Application to the detection of immune complexes and C3 in human sera and of private antigens in cross-reacting parasite extracts.

作者信息

Pontes-de-Carvalho L C, Lannes-Vieira J, Giovanni-de-Simone S, Galvão-Castro B

出版信息

J Immunol Methods. 1986 May 1;89(1):27-35. doi: 10.1016/0022-1759(86)90028-1.

Abstract

An immunoradiometric assay, based on the precipitation of antigen-antibody complexes by polyethylene glycol (PEG) and on the subsequent binding of PEG-soluble radiolabelled staphylococcal protein A to the PEG-insoluble complexes, is described. The assay can be applied to the detection of naturally occurring, circulating immune complexes, and of complexes artificially created by mixing antigen and antibody solutions, which makes it of potential use for the detection of either antigen or antibody in several situations. Pre-treatment of the antibody-containing sera with 3% PEG greatly reduced the background values and increased the sensitivity of the assay. The assay was also applied to the detection and isolation of Leishmania donovani antigens that did not cross-react with antigens of the related parasite Trypanosoma cruzi (private antigens) and private antigens of insect-derived metacyclic trypomastigotes of T. cruzi in relation to culture-derived metacyclic trypomastigotes of T. cruzi. A simple and extremely effective procedure for washing precipitates with just one centrifugation is also described.

摘要

本文描述了一种免疫放射分析方法,该方法基于聚乙二醇(PEG)沉淀抗原-抗体复合物,以及随后将PEG可溶性放射性标记葡萄球菌蛋白A与PEG不溶性复合物结合。该分析方法可用于检测天然存在的循环免疫复合物,以及通过混合抗原和抗体溶液人工形成的复合物,这使其在多种情况下检测抗原或抗体具有潜在用途。用3% PEG预处理含抗体血清可大大降低背景值并提高分析的灵敏度。该分析方法还应用于检测和分离杜氏利什曼原虫抗原,这些抗原与相关寄生虫克氏锥虫的抗原不发生交叉反应(私有抗原),以及克氏锥虫昆虫来源的循环后期锥鞭毛体相对于克氏锥虫培养来源的循环后期锥鞭毛体的私有抗原。本文还描述了一种仅通过一次离心洗涤沉淀物的简单且极其有效的方法。

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