Cell culture medium
The present invention provides a basal cell culture medium and a feed medium with novel amino acid ratios and/or iron choline citrate as iron carrier that result in improved performance of mammalian cell culture processes, such as CHO cultivation and protein production processes, in particular in increased product titer (e.g. of monoclonal antibodies). Also provided are methods for culturing mammalian cells and producing a protein of interest using said basal cell culture medium and optionally feed medium. The invention also provides for a medium platform that comprises (i) the basal cell culture medium and (ii) the feed medium.
1. A medium platform for culturing mammalian cells comprising:
a) a basal cell culture medium for culturing mammalian cells comprising the following amino acids at a molar ratio relative to isoleucine (mM/mM) of:
L-leucine/L-isoleucine of 1.2-2.2,
L-phenylalanine/L-isoleucine of 0.5-0.9,
L-tyrosine/L-isoleucine of 1.5-2.7,
L-threonine/l-isoleucine of 1.0-1.9, and
L-valine/L-isoleucine of 1.0-1.9,
wherein the basal cell culture medium has a total amino acid content of 25 to 150 mM, and
b) a feed medium comprising the following amino acids at a molar ratio relative to isoleucine (mM/mM) of:
L-leucine/L-isoleucine of 2.3-4.2,
L-phenylalanine/L-isoleucine of 0.6-1.1,
L-threonine/l-isoleucine of 1.3-2.4, and
L-valine/L-isoleucine of 1.1-2.0,
wherein the feed medium has a total amino acid content of 100 to 1000 mM.
2. The medium platform of claim 1 , wherein the basal cell culture medium further comprises L-lysine at a molar ratio relative to isoleucine of 1.6-2.9 (mM/mM).
3. The medium platform of claim 1 , wherein the basal cell culture medium further comprises at least one of the following amino acids at a molar ratio relative to isoleucine (mM/mM) of:
L-tryptophan/L-isoleucine of 0.3-0.5,
L-proline/L-isoleucine of 1.6-3.0; or
L-methionine/L-isoleucine of 0.4-0.7.
4. The medium platform of claim 1 , wherein the basal cell culture medium further comprises iron choline citrate at a concentration of 0.1 to 5.0 mM, 0.2 to 2.0 mM, 0.2 to 1.0 mM or 0.4 to 1.0 mM.
5. The medium platform of claim 1 , wherein the feed medium further comprises the following amino acids at a molar ratio relative to isoleucine (mM/mM) of:
L-tyrosine/L-isoleucine of 0.6-1.1, or L-lysine/L-isoleucine of 1.1-2.1, or
L-tyrosine/L-isoleucine of 0.6-1.1, and L-lysine/L-isoleucine of 1.1-2.1.
6. The medium platform of claim 1 , wherein the feed medium further comprises at least one of the following amino acids at a molar ratio relative to isoleucine (mM/mM) of:
L-tryptophan/L-isoleucine of 0.3-0.6,
L-proline/L-isoleucine of 0.9-1.8; or
L-methionine/L-isoleucine of 0.4-0.8.
7. The medium platform of claim 1 , wherein the feed medium additionally comprises iron choline citrate at a concentration of 0.4 to 5 mM, 0.4 to 1.0 mM or 0.5 to 1.0 mM, or 0.5 to 0.6 mM.