A NOVEL COLORIMETRIC ENZYME ASSAY FOR FIBRINOLYSIS

点击翻译按钮获取中文摘要

We have recently developed an assay system for the components of the fibrinolytic pathway. This novel assays sis based on a solid phase enzyme linked fibrin matrix in which plasmin is measured by the release of enzyme from the matrix. We propose to improve certain solid phase facets of the assay in terms of convenience and to extend this technology to produce a series of functional imunoassays for activators and inhibitors: as well as immunoassays for complexes of the activators and inhibitors of this system. The factors of interest include the activators t-PA and urokinase, the plasmin inhibitor, alpha2 - antiplasmin and the activator inhibitor, PAI-1 (plasminogen activator inhibitor -1). The complexes we will study are t-PA -PAI-1 which is indicative of the degree of inhibition at the activation level, and alpha 2 -antiplasmin - plasmin which indicates the extent of activation of the fibrinolytic system. We have arranged to carry out clinical applications of our assay using plasma from three patient populations. The first group is coronary bypass patients who represent a group with serious perturbations of hemostasis, the second group consists of a well characterized patients who have low levels of traditional risk factors for coronary artery disease (CAD) but do in fact have CAD, and finally a group of patients diagnosed as having unstable angina, a group who may be designated "pre myocardial infarction". These groups of patients should allow us to determine the predictive and clinical usefulness of our assays and provide valuable information on the state of activation or inhibition of the fibrinolytic system in these three areas of concern. We also propose use our system as a basic research tool to determine the type of plasminogen activators produced by primary cell cultures of rabbit epithelial cells.