PHYSICAL CHEMISTRY OF THERAPEUTIC PLASMA PROTEINS

该提案描述了对选定的 血浆蛋白，包括抗凝血酶III、Cl-灭活剂、α 2-纤溶酶 抑制剂和纤连蛋白，它们都具有潜在替代价值 治疗患有遗传性或后天性缺陷的患者。 程序 将加速开发新的或改进的隔离方法， 将这些蛋白质以适于临床评价的形式保存。 的 将在聚乙烯存在下评价各种蛋白质的溶解度 乙二醇是一种惰性分级沉淀剂，正在开发作为 用于大规模分馏的乙醇替代品。 我们将寻求 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 将检查蛋白质对各种形式的稳定性 应力，特别强调热稳定性，目标是 确定可对其进行巴氏消毒的条件，以减少 输血性肝炎。 最后，我们建议制定一个广泛的方案， 人类蛋白质-蛋白质和蛋白质-配体相互作用研究 等离子体，重点是使用荧光光谱和高压 液相色谱法 要研究的系统包括：（1）相互作用 在上述三种蛋白酶抑制剂与相关蛋白酶和酶原中， (2)纤连蛋白与纤维蛋白（原）、胶原和肝素的相互作用，（3） 这些蛋白质的不同片段之间的相互作用。 的结果 这些研究除了阐明结构/功能关系外， 应用于荧光结合测定的开发和改进 基于亲和原理的纯化方案。

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.