Methods of producing anti-TNF-alpha antibodies in mammalian cell culture
AssigneeAbbVie Inc.
InventorsItzcoatl A. Pla, Joseph C. Matuck, John C. Fann, Christof Schulz, Nicole A. Roy, David F. Bruton, James McIntire, David Chang Yu-Hsiang, Thomas Seewoester
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Abstract
The invention describes improved methods and compositions for producing a recombinant protein, e.g., an antibody, in mammalian cell culture. In addition, the invention provides improved cell culture media, including improved production media, feed solutions, and combination feeds, which may be used to improve protein productivity in mammalian cell culture.
Claims39
A method of producing an anti-TNFα antibody in a mammalian cell culture, said method comprising:
- a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding the antibody, or a fragment thereof, in a cell culture growth medium to form the mammalian cell culture;
- and
- b) culturing the CHO cells in a cell culture production medium, wherein glucose is added to the mammalian cell culture to maintain a glucose concentration of at least 2 g/L,
- such that the anti-TNFα antibody is produced,
- wherein the anti-TNFα antibody comprises a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:3, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:
- 7, and comprises a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:
- 6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:8.
A method of producing an anti-TNFα antibody in a mammalian cell culture, said method comprising:
- a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding the antibody, or a fragment thereof, in a cell culture growth medium to form the mammalian cell culture;
- and
- b) culturing the CHO cells in a cell culture production medium, wherein glucose is added to the mammalian cell culture to maintain a glucose concentration of at least 2 g/L,
- such that the anti-TNFα antibody is produced,
- wherein the anti-TNFα antibody comprises a light chain variable region (LCVR) comprising the amino acid sequence of SEQ ID NO:1, and a heavy chain variable region comprising the amino acid sequence of SEQ ID NO:2.
A method of producing adalimumab in a mammalian cell culture, said method comprising:
- a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding adalimumab, or a fragment thereof, in a cell culture growth medium to form the mammalian cell culture;
- and
- b) culturing the CHO cells in a cell culture production medium, wherein glucose is added to the mammalian cell culture to maintain a glucose concentration of at least 2 g/L,
- such that adalimumab is produced.
A method of producing an anti-TNFα antibody, or an antigen-binding fragment thereof, in a mammalian cell culture, said method comprising culturing Chinese Hamster Ovary (CHO) cells expressing the anti-TNFα antibody, or the antigen-binding fragment thereof, in a cell culture production medium, wherein glucose is added to the cell culture production medium to maintain a glucose concentration of at least 2 g/L, such that the anti-TNFα antibody, or the antigen-binding fragment thereof, is produced,
- wherein the anti-TNFα antibody, or the antigen-binding fragment thereof, comprises a light chain variable region (LCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:
- 3, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:
- 5, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:
- 7, and comprises a heavy chain variable region (HCVR) having a CDR3 domain comprising the amino acid sequence of SEQ ID NO:
- 4, a CDR2 domain comprising the amino acid sequence of SEQ ID NO:
- 6, and a CDR1 domain comprising the amino acid sequence of SEQ ID NO:
- 8.
Dependencies are inferred from the claim text.