| 摘要 |
<p>The present invention discloses a novel method for the preparation of recombinant human proteins expressed in human cells. Specifically, the present invention relates to novel methods for the preparation of human recombinant thrombin and human recombinant fibrinogen. Moreover, the method employs serum-free culturing conditions and therefore provides recombinant human proteins expressed in human cells of increased safety to the patient when used in human medical treatments. In addition, the immunogenic response to the recombinant human proteins expressed in human cells may be lower. Human recombinant thrombin is expressed in the human embryonic kidney 293 cell line and the protein can be prepared using two different routes, one starting from a point mutated prothrombin with gla and Kringle 1 and 2 domains, and maintaining these domains during the process; the other one starting with prothrombin (non-mutated), via a prethrombin with a HPC4-Kringle 2 domain and subjecting this prethrombin to a point mutation. Human recombinant fibrinogen is expressed in the human embryonic kidney 293 cell line.</p> |
