Cell culture compositions and methods for polypeptide production
AssigneeGenentech, Inc.
InventorsNatarajan VIJAYASANKARAN, Steven J. MEIER, Melissa S. MUN, Sharat VARMA, Yi YANG, Boyan ZHANG, Silvana R. AREVALO, Martin GAWLITZEK, Veronica CARVALHAL
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Abstract
Cell culture media, such as chemically defined cell culture media, are provided, as are methods of using the media for cell growth (i.e., cell culture) and polypeptide (e.g., antibody) production. Compositions comprising polypeptides produced by the methods are also provided.
Claims35
A method of culturing cells, comprising the step of contacting the cells with a cell culture medium comprising:
- from about 300 mg/L to about 1200 mg/L cystine;
- from about 0.05 mg/L to about 1.0 mg/L vitamin B2;
- from about 0.05 mg/L to about 10.0 mg/L vitamin B6;
- from about 0.05 mg/L to about 12.0 mg/L vitamin B9;
- and
- from about 0.05 to about 2.5 mg/L vitamin B12.
A method of producing a polypeptide comprising the step of culturing in a cell culture medium a cell comprising an isolated nucleic acid encoding the polypeptide, wherein:
- (a) the cell culture medium comprises:
- from about 300 mg/L to about 1200 mg/L cystine;
- from about 0.05 mg/L to about 1.0 mg/L vitamin B2;
- from about 0.05 mg/L to about 10.0 mg/L vitamin B6;
- from about 0.05 mg/L to about 12.0 mg/L vitamin B9;
- from about 0.05 to about 2.5 mg/L vitamin B12;
- and
- (b) the cell expresses the polypeptide.
17-30. (canceled)
A kit for supplementing a cell culture medium with chemically defined constituents, the kit comprising:
- cystine in an amount to provide from about 300 mg/L to about 1200 mg/L cystine in the cell culture medium;
- vitamin B2 in an amount to provide from about 0.05 mg/L to about 1.0 mg/L vitamin B2 in the cell culture medium;
- vitamin B6 in an amount to provide from about 0.05 mg/L to about 10.0 mg/L vitamin B6 in the cell culture medium;
- vitamin B9 in an amount to provide from about 0.05 mg/L to about 12.0 mg/L vitamin B9 in the cell culture medium;
- and
- vitamin B12 in an amount to provide from about 0.05 to about 2.5 mg/L vitamin B12.
34-39. (canceled)
A cell culture medium comprising:
- from about 300 mg/L to about 1200 mg/L cystine;
- from about 0.05 mg/L to about 1.0 mg/L vitamin B2;
- from about 0.05 mg/L to about 10.0 mg/L vitamin B6;
- from about 0.05 mg/L to about 12.0 mg/L vitamin B9;
- and
- from about 0.05 to about 2.5 mg/L vitamin B12.
41-49. (canceled)
A method of culturing cells, comprising the step of contacting the cells with a cell culture medium comprising:
- from about 300 mg/L to about 1200 mg/L cystine;
- from about 2 μM to about 80 μM ferric citrate;
- and
- from about 0.05 μM to about 0.5 μM hydrocortisone.
51-59. (canceled)
A method of producing a polypeptide comprising the step of culturing in a cell culture medium a cell comprising an isolated nucleic acid encoding the polypeptide, wherein:
- (a) the cell culture medium comprises:
- from about 300 mg/L to about 1200 mg/L cystine;
- from about 2 μM to about 80 μM ferric citrate;
- and
- from about 0.05 μM to about 0.5 μM hydrocortisone;
- and
- (b) the cell expresses the polypeptide.
61-71. (canceled)
A kit for supplementing a cell culture medium with chemically defined constituents, the kit comprising:
- cystine in an amount to provide from about 300 mg/L to about 1200 mg/L cystine in the cell culture medium;
- ferric citrate in an amount to provide a concentration of from about 2 μM to about 80 μM ferric citrate in the cell culture medium;
- and
- hydrocortisone in an amount to provide a concentration of from about 0.05 μM to about 0.5 μM hydrocortisone in the cell culture medium.
75-79. (canceled)
A cell culture medium comprising:
- from about 300 mg/L to about 1200 mg/L cystine;
- from about 2 μM to about 80 μM ferric citrate;
- and
- from about 0.05 μM to about 0.5 μM hydrocortisone.
81-85. (canceled)
Dependencies are inferred from the claim text.