Affinity Chromatographic Purification of Plasminogen Activators from Mammalian Cell Culture Broth

Plasminogen Activators (PAs) are serine proteases that convert the plasminogen into plasmin. They are used for the treatment of thromboembolic disorders since their physiological function is to dissolve fibrin clots. The major PAs in use are characterized by low potency, side effects, low bioavailability, immunogenicity and high costs. The objective of this work is to isolate novel PAs with higher fibrin-specificity from mammalian cell culture broth. A new PA has been isolated from HEK-293 cell culture broth through a multi-step isolation and purification process. These plasminogen activators were isolated using ammonium sulfate precipitation process and a commercially available matrix: lysine Sepharose 4B in a sequential process. The final PA product was further characterized with respect to protein concentration, enzyme activity, and electrophoresis.