NOVEL ACTIVITY AND IMMUNO - ASSAYS FOR PAI-1

纤溶酶原激活受多种抑制物的调节。他禁食 T-PA和u-PA的作用抑制物被指定为纤溶酶原 激活物抑制物-1)PAI-1)。检测到这一水平的升高 急性期蛋白提示止血作用失衡 系统。为了全面评估PAI-1血浆水平，有必要 用抗原法和活性测定法测定蛋白质。我们提出了一个 一种允许测量总血浆抗原的方法 水平、PAI-1/t-PA抗原水平、PAI-1活性水平和 玻璃连蛋白/纤溶酶原激活物-1复合体的活性和抗原水平 代表PAI-1的一个特殊、稳定的亚群，它可能具有 抗凝血酶活性。活性测定涉及一种酶的使用 标记的纤维蛋白基质作为t-PA的底物；剩余活性将 允许测量PAI-1抑制活性。我们建议使用 标准酶联免疫吸附试验和超灵敏的酶联免疫吸附试验 技术。这是因为游离PAI-1抗原和复合体的水平 PAI-1至少相差10倍。随着试剂的制备 进行这些化验，我们将能够对以下化验进行初步测试 血浆成分，如玻璃凝集素/PAI-1复合体和u-PA PAI-1 复合体。

Plasminogen activation is modulated by a number of inhibitors. he fast acting inhibitor of t-PA and u-PA has been designated plasminogen activator inhibitor-1 )PAI-1). The detection of raised levels of this acute phase protein is indicative of an imbalance in the hemostatic system. In order to fully assess PAI-1 plasma levels it is necessary to measure the protein by both antigenic and activity assays. We propose an approach which will allow the measurement of total plasma antigen levels, PAI-1/t-PA antigen levels, PAI-1 activity levels, and the activity and antigen levels of vitronectin/PAI-1 complex which represents a special, stable subpopulation of PAI-1 which may have antithrombin activity. The activity assay involves the use of an enzyme labelled fibrin matrix as the substrate for t-PA; residual activity will allow the measurement of PAI-1 inhibitory activity. We propose to use both standard ELISA assay and the ultrasensitive ELISA-ELCA assay technique. This is because levels of free PAI-1 antigen and complexed PAI-1 differ by at least 10 fold. With the preparation of reagents to carry out these assays we will be able to do initial tests on assays for plasma components such as the vitronectin/PAI-1 complexes and u-PA PAI-1 complexes.