NOVEL ACTIVITY AND IMMUNO - ASSAYS FOR PAI-1

纤溶酶原激活由许多抑制剂调节。他快 t-PA和u-PA的作用抑制剂已被指定为纤溶酶原 激活剂抑制剂-1）派-1）。检测到高水平的这种 急性时相蛋白指示止血剂中的不平衡 系统为了全面评估派-1血浆水平，有必要 通过抗原性和活性测定来测量蛋白质。我们提出了一个 允许测量总血浆抗原的方法 派-1/t-PA抗原水平，派-1活性水平， 玻连蛋白/派-1复合物的活性和抗原水平， 代表派-1的一个特殊的、稳定的亚群， 抗凝血酶活性活性测定涉及酶的使用 标记的纤维蛋白基质作为t-PA的底物;残余活性将 允许测量派-1抑制活性。我们建议使用 标准ELISA测定和超灵敏ELISA-ELCA测定 法这是因为游离派-1抗原和复合的PAI-1抗原的水平 派-1至少相差10倍。随着试剂的制备， 进行这些检测，我们将能够对检测进行初步测试， 血浆组分如玻连蛋白/派-1复合物和u-PA派-1 配合物

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.