| 摘要 |
601860 Disclosed is a method of producing a polypeptide in a large-scale production cell culture comprising the steps of: providing a cell culture comprising; mammalian cells that contain a gene encoding a polypeptide of interest, which gene is expressed under condition of cell culture; and a medium containing glutamine, having a cumulative amino acid amount per unit volume greater than about 70 mM, a molar cumulative glutamine to cumulative asparagine ratio of less than about 2, and having a medium characteristic selected from the group consisting of (i) a molar cumulative glutamine to cumulative total amino acid ratio ofless than about 0.2, (ii) a molar cumulative inorganic ion to cumulative total amino acid ratio between about 0.4 to 1, and (iii) a combined cumulative amount of glutamine and asparagine per unit volume of greater than about 16 mM, and combinations thereof; wherein the cumulative total amount of histidine, isoleucine, leucine, methionine, phenylalanine, tryptophan, tyrosine, and proline per unit volume in said medium is greater than approximately 25 mM; maintaining said culture in an initial growth phase under a first set of culture conditions for a first period of time sufficient to allow said cells to reproduce to a viable cell density within a range of about 20%-80% of the maximal possible viable cell density if said culture were maintained under the first set of culture conditions; changing at least one of the culture conditions, so that a second set of culture conditions is applied; maintaining said culture for a second period of time under the second set of conditions and for a second period of time so that the polypeptide accumulates in the cell culture, provided that the polypeptide is not a TNFR-Fc, TNFR-Ig or an antibody to a ABeta fragment of amyloid precursor protein. |
