METHOD FOR MASS PRODUCING HUMAN BLOOD COAGULATION FACTOR VII DERIVATIVE,申请号EP20140839952-传众专利搜索

METHOD FOR MASS PRODUCING HUMAN BLOOD COAGULATION FACTOR VII DERIVATIVE

申请公布号：EP3040419(A1)

申请号：EP20140839952

申请日期：2014.08.27

申请公布日期：2016.07.06

申请人：

HANMI PHARM. CO., LTD.

发明人：KANG, HEE CHUL;KIM, JIN YOUNG;LEE, BYUNG SUN;KIM, HYUN UK;CHOI, IN YOUNG;KWON, SE CHANG

分类号：C12N15/12;C07K14/745;C12N9/06;C12N9/64;C12N15/85;C12P21/02

主分类号：C12N15/12

摘要：The present invention provides a method for mass production of human coagulation factor VII derivatives, containing: a) constructing an expression vector, containing i) a nucleotide sequence of a dihydrofolate reductase promoter devoid of at least one CCGCCC repeat sequence from the GC-rich region thereof and a nucleotide sequence encoding a dihydrofolate reductase (DHFR) operably linked thereto, and ii) a nucleotide sequence of a cytomegalovirus (CMV) early gene promoter and a nucleotide sequence encoding a human coagulation factor VII derivative operably linked thereto; b) transfecting an animal cell line with the expression vector of step a); c) culturing the transfected animal cell line of step b) in the presence of a dihydrofolate reductase inhibitor to select a cell line capable of expressing the human coagulation factor VII derivative with high efficiency; and d) culturing the animal cell line selected from step c) by adding at least one selected from the group consisting of sodium butyrate, vitamin K, and a culture medium supplement. Employing a vector containing a DHFR promoter devoid of GC-rich repeat sequences, the present invention can express human coagulation factor VII derivates with high efficiency on a large scale, and thus can be useful for the preparation of therapeutic agents for hemophilia.