Köttgen E, Hoeft S, Müller C, Hell B
J Clin Chem Clin Biochem. 1986 Aug;24(8):541-9. doi: 10.1515/cclm.1986.24.8.541.
So far, soluble fibronectin has been quantitated mostly by immunological techniques. In this investigation we show that an immunological assay provides reliable results only with intact fibronectin. Fibronectin fragments resulting from proteolysis give rise to falsely raised values. We present four functional tests based on the sandwich (ELISA) technique on microtitre plates. These quantify fibronectin on the basis of its binding capacity to collagen, heparin, fibrin and carboxy-group-modified IgG with high sensitivity, specificity and precision. Analysis of the bioactivity spectrum of intact fibronectin is not disturbed by fibronectin fragments. Furthermore we demonstrate interferences, in particular between heparin and collagen in their mutual binding to fibronectin. This provides new indications of a "substrate activation" of fibronectin.
到目前为止，可溶性纤连蛋白大多通过免疫技术进行定量。在本研究中，我们表明免疫测定仅对完整的纤连蛋白提供可靠结果。蛋白水解产生的纤连蛋白片段会导致值假性升高。我们在微量滴定板上基于夹心（ELISA）技术提出了四种功能测试。这些测试基于纤连蛋白与胶原蛋白、肝素、纤维蛋白和羧基修饰的IgG的结合能力，以高灵敏度、特异性和精密度对纤连蛋白进行定量。完整纤连蛋白的生物活性谱分析不受纤连蛋白片段的干扰。此外，我们证明了干扰，特别是肝素和胶原蛋白在它们与纤连蛋白相互结合方面的干扰。这为纤连蛋白的“底物激活”提供了新的迹象。
