PHYSICAL CHEMISTRY OF THERAPEUTIC PLASMA PROTEINS

这项建议描述了对选定的 血浆蛋白包括抗凝血酶III、氯灭活物、α2-纤溶酶 抑制剂和纤维连接蛋白，所有这些都有潜在的替代价值 遗传性或获得性缺陷患者的治疗。该计划 将加快开发新的或改进的分离和 以适合临床评估的形式保存这些蛋白质。这个 在聚乙烯存在的情况下，将评估各种蛋白质的溶解度。 乙二醇是一种惰性分级沉淀剂，正在作为一种 乙醇的替代品，用于大规模分馏。我们将想方设法 选择性地操纵特定蛋白质的溶解度，例如通过 添加它们优先与之相互作用的物质。其次，这些 我们将检查蛋白质对各种形式的稳定性。 应激，特别强调热稳定性，目标是 定义可对其进行巴氏灭菌以降低风险的条件 输血性肝炎。最后，我们建议制定一项广泛的计划 人体蛋白质-蛋白质和蛋白质-配体相互作用的研究 等离子体，重点是荧光光谱和高压的使用 液体色谱学。要调查的系统包括：(1)相互作用 在上述三种具有相关酶和酵素的蛋白酶抑制剂中， (2)纤维连接蛋白与纤维蛋白原、胶原和肝素的相互作用； 这些蛋白质不同片段之间的相互作用。结果是 这些研究，除了澄清结构/功能关系外，还将 应用于荧光结合分析的发展及改进 基于亲和力原理的纯化方案。

This proposal describes research into the physicochemical properties of selected plasma proteins including antithrombin III, Cl-inactivator, alpha2-plasmin inhibitor, and fibronectin, all of which have potential value for replacement therapy in patients with either inherited or acquired deficiences. The program will expedite the development of new or improved methods of isolating and preserving these proteins in a form suitable for clinical evaluation. The solubility of various proteins will be evaluated in the presence of polyethylene glycol, an inert fractional precipitating agent, which is being developed as an alternative to ethanol for large-scale fractionation. We will seek ways of selectively manipulating the solubility of specific proteins, for example by addition of substances with which they preferentially interact. Secondly, these proteins will be examined with respect to their stability towards various forms of stress, with particular emphasis on thermal stability, the objective being to define conditions under which they can be pasteurized to diminish the risk of transfusion hepatitis. Finally, we propose to develop a broad program of investigation of protein-protein and protein-ligand interactions in human plasma, with emphasis on the use of fluorescence spectroscopy and high pressure liquid chromatography. Systems to be investigated include: (1) the interaction of the above three protease inhibitors with the relevant proteases and zymogens, (2) the interaction of fibronectin with fibrin(ogen), collagen, and heparin, (3) the interaction between various fragments of these proteins. The results of these studies, in addition to clarifying structure/function relationships, will be applied to the development of fluorescent binding assays and improved purification schemes based on affinity principles.