| CPC C12N 5/0018 (2013.01) [C12P 21/00 (2013.01); C12N 2500/40 (2013.01); C12N 2500/60 (2013.01)] | 16 Claims |

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1. A method of producing a polypeptide of interest in a glutamine-free cell culture medium, the method comprising:
culturing cells expressing the polypeptide of interest in the glutamine-free cell culture medium under conditions that support expression of the polypeptide of interest,
wherein the glutamine-free cell culture medium comprises: (i) between about 20 μM and about 100 μM hypoxanthine; (ii) between about 15 μM and about 20 μM thymidine; and (iii) adenosine and/or at least one corticosteroid, and
wherein the cells are initially maintained in a culture containing glutamine and are transitioned to the glutamine-free cell culture medium for production phase.
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