abstract |
PURPOSE: To readily obtain a human urinary plasminogen activator derivative useful for medicines, etc., for prevention of thrombus by chemically modifying native single chain urokinase in a growth factor-like domain(GFD). n CONSTITUTION: Urokinase (uPA) is collected from a human genome library. Then, the uPA gene is inserted into an expression vector such as RSV-neo vector of retrovirus (RSV) to modify the plasmid vector and GFD of uPA gene is changed. The resultant plasmid vector RSVuPA is introduced into an animal cell to provide a transformed cell. Then, the cell is cultured in Dulbecco-modified Eagle medium, etc., containing 10-20wt.% inactive fetal bovine serum, etc. The resultant cultured cell is selected by immunoamidolytic assay to provide glucosylated plasminogen activator (GASGA)-producing clone. Then, the clone is cultured in a culture medium containing aprotinin, etc., to produce human urine plasminogen activator derivative. n COPYRIGHT: (C)1989,JPO |