公开(公告)号：JP2014193187A

公开(公告)日：2014-10-09

申请号：JP2014136448

申请日：2014-07-02

申请人： Roche Diagnostics Gmbh ,  ロシュ ダイアグノスティックス ゲーエムベーハー

发明人： BRANDT MICHAEL ,  FRANZE REINHARD ,  PESSARA ULRICH

IPC分类号： C12Q1/02 ,  A61K38/00 ,  C07K14/505 ,  C12N5/08 ,  C12N15/09 ,  C12N15/10 ,  C12N15/62 ,  C12P21/02

CPC分类号： C07K14/505 ,  A61K38/00 ,  C07K2319/02 ,  C07K2319/036 ,  C07K2319/75 ,  C12N15/10 ,  C12N15/625

摘要： PROBLEM TO BE SOLVED: To provide a method for the selection of human cells for the production of human proteins by endogenous gene activation in order to produce human proteins in economical yields and in a form which is suitable for the production of a pharmaceutical preparation.SOLUTION: A method for the selection of human cell lines for the preparation of human proteins by the activation of a target gene endogenously present in a cell is characterized in that (a) a human cell line is tested for the presence of the following features: (i) the target gene with a desired nucleic acid sequence; (ii) at least 5 population doublings of the cell number within 14 days in a suspension culture; and (iii) at least 5 population doublings of the cell number within 14 days in a serum-free culture medium, and (b) using a cell line having features (i), (ii), and (iii) as starting cell line for the endogenous activation of the target gene.

摘要翻译： 要解决的问题：提供一种通过内源基因激活选择人细胞用于生产人类蛋白质的方法，以便以经济的产率和以适于制备药物制剂的形式产生人蛋白质。 ：通过活化内源存在于细胞中的靶基因来选择用于制备人蛋白质的人细胞系的方法的特征在于（a）测试人细胞系存在以下特征：（ i）具有所需核酸序列的靶基因; （ii）在悬浮培养中14天内至少5次细胞数量的倍增; 和（iii）在无血清培养基中在14天内至少5次细胞数量的倍增，和（b）使用具有特征（i），（ii）和（iii）的细胞系作为起始细胞系 用于靶基因的内源性激活。