Abstract:
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.
Abstract:
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.
Abstract:
PROBLEM TO BE SOLVED: To provide a method for producing a glycosylated polypeptide. SOLUTION: The method for producing the appropriately glycosylated polypeptide by culturing eukaryotic cells and isolating the resultant polypeptide from the medium and/or the cells is provided. By this method, the desired glycosylated polypeptide can be produced by recombination using endogenous gene activation or by the cells spontaneously. A method for obtaining a highly glycosylated desired protein such as EPO in high yield by adding a mixture of at least two carbohydrates to a medium having eukaryotic cells is also provided. COPYRIGHT: (C)2003,JPO