AbstractPlasminogen activators of both urokinase‐type (u‐PA) and tissue‐type (t‐PA) have been found in conditioned medium of cultured human fibroblasts. To purify t‐PA, the total PA activity from hectoliter quantities of conditioned medium was first concentrated with about 10‐fold increase in specific activity via zinc chelate‐sepharose affinity chromatography. Three different methods were employed in an attempt to separate t‐PA from u‐PA: (a) concanavalin A‐sepharose affinity chromatography; (b) fibrin‐celite affinity chromatography; and (c) immunoaffinity chromatography using anti‐tPA monoclonal antibody immobilized on sepharose. Of the three methods, only the monoclonal antibody procedure effected complete separation of the two enzymes. t‐PA was subsequently purified to homogeneity using size exclusion, high performance liquid chromatography followed by p‐aminobenzamidine‐ag
展开▼
