免疫复合物中抗原的检测。

The detection of antigens in immune complexes.

作者信息

Heimer R, Glick D L, Abruzzo J L

出版信息

Scand J Immunol. 1981;13(5):441-6. doi: 10.1111/j.1365-3083.1981.tb00154.x.

DOI:10.1111/j.1365-3083.1981.tb00154.x

Abstract

A procedure is described for the purification of soluble immune complexes (IC) from biologica fluids and for the detection of antibody-bound antigen. A model IC, prepared with various amounts of human serum albumin (HSA) and constant amounts of anti-HSA, gave a 44% recovery after gel filtration on Sephacryl S-300, affinity chromatography on protein A--Sepharose, and concentration of eluates there from before analysis by sodium dodecyl sulphate--polyacrylamide gel electrophoresis. It was calculated that 20 ng of antibody-bound HSA should be detectable when polyacrylamide gels are subjected to a direct radioimmunoassay involving the use of rabbit anti-HSA, 125I-protein A, and autoradiography, thus defining the sensitivity of the procedure for detecting IC-bound antigen in a model system. The procedure has direct relevance to the examination of IC of unknown composition present in sera of individuals with various diseases.

摘要

本文描述了一种从生物体液中纯化可溶性免疫复合物（IC）以及检测抗体结合抗原的方法。用不同量的人血清白蛋白（HSA）和恒定剂量的抗HSA制备的模型IC，在经Sephacryl S - 300凝胶过滤、蛋白A - 琼脂糖亲和层析以及在通过十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳分析之前对洗脱液进行浓缩后，回收率为44%。据计算，当聚丙烯酰胺凝胶进行直接放射免疫测定（使用兔抗HSA、¹²⁵I - 蛋白A和放射自显影）时，应能检测到20 ng的抗体结合HSA，从而确定了该方法在模型系统中检测IC结合抗原的灵敏度。该方法与检测患有各种疾病个体血清中未知组成的IC直接相关。