A NOVEL COLORIMETRIC ENZYME ASSAY FOR FIBRINOLYSIS

一种新型的纤维蛋白溶解比色酶测定方法

• 批准号: 3508681
• 负责人: MARK X TRISCOTT
• 金额: $ 30.28万
• 依托单位: AMPLISTAR, INC.
• 依托单位国家: 美国
• 财政年份: 1989
• 资助国家: 美国
• 起止时间: 1989-09-30 至 1991-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/3508681
• 关键词: angina pectoris; antifibrinolytic agents; bioassay; blood coagulation tests; diagnosis design /evaluation; diagnostic catheterization; disease /disorder proneness /risk; enzyme linked immunosorbent assay; enzyme mechanism; enzyme substrate; extracellular matrix; fibrin; fibrinogen; fibrinolysis; fibrinolytic therapy; hemostasis; human subject; human tissue; immunologic techniques; myocardial infarction; plasmin; plasminogen; plasminogen activator; rapid diagnosis; streptokinase; urokinase

Thrombolytic therapy has become an increasingly important tool in the treatment of acute myocardial infarction. The most recently approved therapeutic agent for this purpose is t-PA (tissue plasminogen activator) which has the advantages of being fibrin specific and is rapidly cleared in vivo by the liver. The detection and quantitation of this component of the fibrinolytic system as well as its inhibitors is still time consuming and is measured by lysis zone areas. We propose a new technique for the quantitation of all the components of the fibrinolytic system including t-PA, plasminogen, plasmin, alpha 2-antiplasmin, and antiactivators (eg PAI-1). The assay uses a solid phase enzyme labelled fibrin as a substrate for locally generated plasmin. The manipulation of concentrations of activators and inhibitors will directly influence the amount of plasmin produced. Quantitation of the resulting plasmin is achieved through the detection and measurement of enzyme labelled fibrin fragments released through the proteolytic action of the plasmin. This assay technique has a number of advantages over existing assays including the use of solid phase fibrin as a substrate and rate enhancer which parallels the situation in vivo. The assay is also convenient for handling multiple samples, lends itself to automation and because of its sensitivity is extremely cost effective.

溶栓治疗已成为一种日益重要的工具 治疗急性心肌梗死。最多的 最近批准的用于此目的的治疗剂是t-PA (组织纤溶酶原激活剂)，其优点是 纤维蛋白是特异的，并在体内被肝脏迅速清除。这个 纤溶酶组分的检测和定量 系统及其抑制剂仍然很耗时，而且 以裂解区面积衡量。我们提出了一种新的技术，用于 纤溶系统所有成分的定量测定 包括t-PA、纤溶酶原、纤溶酶、α2-抗纤溶酶和 抗激活剂(如PAI-1)。该检测使用了一种固相酶 标记的纤维蛋白作为局部产生的纤溶酶的底物。这个 控制激活剂和抑制剂的浓度将 直接影响纤溶酶的产生量。量化 所产生的纤溶酶是通过检测和 酶标记纤维蛋白片段释放量的测定 通过纤溶酶的蛋白分解作用。这个化验 与现有的检测方法相比，这项技术有许多优点 包括使用固相纤维蛋白作为底物和比率 与体内情况类似的增强剂。化验结果也是 便于处理多个样品，适合于 自动化，因为它的敏感性是极其昂贵的 有效。