TBBP_Plasminogen^J Plasmin

hemostasis requires interaction of platelets, coagulation and fibrinolytic factors, endothelium, pro- and anti-inflammatory mediators, leukocytes. Clot formation normally initiated by vascular injury (platelet plug reinforced by vascular injury via extrinsic pathway), anticoagulants (AT-III, protein C) localize thrombosis to sites of vascular injury, clot formation balanced by plasmin-mediated fibrinolysis = formation of D-dimers and other fibrin degradation products.

extrinsic pathway: fibrin clot formation in response to tissue injury. Intrinsic pathway: fibrin clot formation in response to an abnormal vessel wall (f.e. by contact with lipoprotein particles or bacteria)

activation plasminogen to plasmin with tPA by cleavage of N-terminal peptide (nicking Arg 561 at activation side), plasmin degrades fibrin clot unless it gets repressed by Alpha2-Antiplasmin (control)

zymogen, inactive form of the enzyme and conversion to plasmin, single chain glycoprotein, 92 kD, 0.2 g/L, synthesis in liver

serine protease (homology to trypsin), 24 disulfide bridges, nonspecific protease, degrades fibrin clots to maintain clot hemostasis, plasmin is rapidly inactivated

purify plasminogen, then activate using a plasmin activator (tPA). Affinity chr. Leveraging the kringle regions. Critical to stabilize plasmin under conditions to avoid autodegradation, viral inactivation must be present. 1. Purification of plasminogen (DF/viral reduction --> AFC --> freeze. 2. purification of plasmin and removal of impurities (viral inactivation, AFC, hydrophobic chr., UF/DF, nanofiltration), 3. formulation, fill and freeze-dry