Enrico Di Cera, M.D., Nicola Pozzi, Ph.D., Sergio Barranco-Medina, Ph.D.
This invention discloses a novel technology for the manufacture of biopharmaceutics and specifically recombinant serine proteases. Engineering of the activation domain generates a precursor that auto-activates in aqueous buffer to form the mature active enzyme. Illustrative active enzymes include serine proteases such as wild type thrombin and the anticoagulant thrombin mutant WE for therapeutic use. This technology would eliminate the need of exogenous activators such as the snake venom ecarin or the prothrombinase complex, therefore improving the safety, quality and cost of the final product. IP includes an optimized scaled-up protocol for the production of the auto-activating thrombin anticoagulant WE.
Recombinant DNA plasmids comprising prothrombin, prethrombin-1 and prethrombin-2.