INTERACTION OF FIBRONECTIN WITH ANTIBODIES AND COLLAGEN IN RADIOIMMUNOASSAY
INTERACTION OF FIBRONECTIN WITH ANTIBODIES AND COLLAGEN IN RADIOIMMUNOASSAY
复制标题
DOI：
10.1016/0005-2795(78)90003-x
复制
发表时间：
1978-01-01
期刊：
影响因子：
--
通讯作者：
ENGVALL, E
中科院分区：
文献类型：
--
作者：
RUOSLAHTI, E;VUENTO, M;ENGVALL, E
将明胶纯化的小鼠和人纤维连接蛋白进行放射性碘标记，得到的标记蛋白中60-70%的放射性可以与抗体过剩的抗纤维连接蛋白结合。用其他方法分离的纤维连接蛋白制剂标记后免疫反应性较低。标记的纤维连接蛋白的主要部分保留了它与胶原的亲和力。这使得可以通过在明胶-琼脂糖上分离碘化蛋白来去除弱免疫反应物质。标记蛋白的结合部分和洗脱部分与抗体和胶原蛋白的结合率均在90%以上。研究了含有纤维连接蛋白的样品中可能存在的胶原蛋白对纤维连接蛋白放射免疫分析的影响以及抗纤维连接蛋白对纤维连接蛋白-胶原相互作用的影响。用放射免疫法测定纤维连接蛋白的含量不受样品中胶原的存在的影响，表明纤维连接蛋白与抗体的相互作用不受胶原的存在的影响。当标记蛋白与纯化的纤维连接蛋白抗体混合时，标记的纤维连接蛋白与胶原的结合被抑制。当抗体通过明胶柱时，先前与明胶柱结合的纤维连接蛋白被释放。抗体与纤维连接蛋白相互作用的亲和力明显高于纤维连接蛋白与胶原的亲和力。
Radioiodination of the gelatin-purified mouse and human fibronectins gave labeled proteins from which 60-70% of the radioactivity could be bound to anti-fibronectin in antibody excess. Fibronectin preparations isolated using other methods showed lower immunoreactivity after labeling. A major part of the labeled fibronectin retained its affinity for collagen. This allowed removal of weakly immunoreactive material by fractionation of the iodinated protein on gelatin-Sepharose. The bound and eluted fraction of the labeled protein showed more than 90% binding to antibody and to collagen. The effect of collagen, a component likely to be present in samples containing fibronectin, on fibronectin radioimmunoassay [RIA] and the effect of anti-fibronectin on the fibronectin-collagen interaction were studied. Quantitation of fibronectin by RIA was found to be unaffected by the presence of collagen in the sample, suggesting that the interaction of fibronectin with antibody was not affected by the presence of collagen. The binding of labeled fibronectin to collagen was inhibited when the labeled protein was mixed with purified antibodies to fibronectin. Fibronectin previously bound to a gelatin column was released when antibodies were passed through the column. The avidity of the antibody-fibronectin interaction apparently is higher than that between fibronectin and collagen.