CELL LINE COEXPRESSING HEAVY AND LIGHT CHAINS OF BLOOD COAGULANT FACTOR VIII AND VON WILLEBRAND FACTOR AND A PROCESS FOR PREPARING A RECOMBINANT FACTOR VIII BY EMPOLYING THE SAME

摘要

PURPOSE: Provided is a cell line expressing light chain and heavy chain of blood coagulation Factor VIII and von Willebrand factor simultaneously. And a preparation method of recombinant blood coagulation Factor VIII is also provided. CONSTITUTION: A preparation method of recombinant blood coagulation Factor VIII is composed of the following steps of: (a) synthesizing recombinant Chinese Hamster Ovary cell line A30 expressing heavy strain and light strain of blood coagulation Factor VIII simultaneously by cotransfection of Chinese Hamster Ovary cell line with heavy chain strain expression factor of human blood coagulation Factor VIII, pCMVFviii-A and light strain expression vector pCMVFviii-C; (b) synthesizing recombinant CHO cell line V88 (KCTC 0421BP) expressing heavy strain, light strain of blood coagulation Factor VIII and von Willebrand factor simultaneously by transfection of the recombinant CHO cell line A30 with expression vector of Von Willebrand factor pCMV-vWF; (c) adapting the recombinant CHO cell line A30 or recombinant CHO cell line V88(KCTC 0421BP) to methotrexate, to amplify a target cell; and (d) cultivating the adapted recombinant CHO cell line A30 or recombinant CHO cell line V88 (KCTC 0421BP) in a serum free medium or medium including serum, to obtain human blood coagulation Factor VIII.