CELL CULTURE IMPROVEMENTS
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.
1 . A method of producing an antibody in a mammalian cell culture such that the titer of the antibody is at least 100 mg/L, said method comprising
a) culturing mammalian cells comprising a nucleic acid encoding the antibody in a cell culture production medium;
b) adding sodium butyrate, N-acetylcysteine, or a combination thereof, to the cell culture medium, wherein the sodium butyrate is added to a final concentration of about 0.1 mM to 10 mM and the N-acetylcysteine is added to a final concentration of about 1 mM to 80 mM,
such that the antibody is produced at a titer of at least 100 mg/L.
2 . The method of claim 1 , wherein the antibody titer is at least 150 mg/L.
3 . A method of producing an antibody in a mammalian cell culture such that the titer of the antibody is at least 10% greater than a control mammalian cell culture, said method comprising
a) culturing mammalian cells comprising a nucleic acid encoding the antibody in a cell culture production medium; and
b) adding sodium butyrate, N-acetylcysteine, or a combination thereof, to the cell culture medium, wherein the sodium butyrate is added to a final concentration of about 0.1 mM to 10 mM and the N-acetylcysteine is added to a final concentration of about 1 mM to 80 mM,
such that the titer of the antibody is at least 10% greater than the control, wherein the control mammalian cell culture comprises step a) and excludes step b).
4 . The method of claim 3 , wherein the antibody titer of the mammalian cell culture is improved at least 29% over the control mammalian cell culture.
5 . The method of claim 3 , wherein the antibody titer of the mammalian cell culture is improved at least 40% over the control mammalian cell culture.
6 . The method of claim 3 , wherein the antibody titer of the mammalian cell culture is improved at least 70% over the control mammalian cell culture.
7 . The method of claim 3 , wherein the antibody titer of the mammalian cell culture is at least 90% greater than the control mammalian cell culture.
8 . The method of claim 1 , wherein sodium butyrate, N-acetylcysteine, or combination thereof, is added to the mammalian cell culture during the growth phase of the mammalian cell culture.
9 . The method of claim 1 , wherein the sodium butyrate, N-acetylcysteine, or combination thereof is added to the mammalian cell culture between days 4 and 7 of the culture time.
10 . The method of claim 1 , wherein the sodium butyrate, N-acetylcysteine, or combination thereof is added to the mammalian cell culture on day 0 of the culture time.
11 . The method of claim 1 , wherein the final concentration of sodium butyrate is about 0.1 mM to 10 mM.
12 . The method of claim 1 , wherein the final concentration of sodium butyrate is about 0.1 mM to 8.0 mM.
13 . The method of claim 1 , wherein the final concentration of N-acetylcysteine is about 20 mM to 60 mM.
14 . The method of claim 1 , wherein the final concentration of N-acetylcysteine is about 10 mM.
15 . The method of any claim 1 , wherein the final concentration of N-acetylcysteine is about 8 mM.
16 . A method of extending longevity of a mammalian cell culture by at least 35% in comparison to a control mammalian cell culture, said method comprising
a) culturing mammalian cells comprising a nucleic acid encoding the antibody in a cell culture production medium; and
b) adding about 1 mM to 80 mM N-acetylcysteine to the cell medium; a
such that the longevity of the mammalian cell culture is extended by at least 35% in comparison to a control mammalian cell culture, wherein the control mammalian cell culture comprises step a) and excludes step b).
17 . The method of claim 16 , wherein the longevity of the mammalian cell culture is extended at least about 45% in comparison to the control mammalian cell culture.
18 . The method of claim 16 , wherein the longevity of the mammalian cell culture is extended at least about 55% in comparison to the control mammalian cell culture.
19 . The method of claim 16 , comprising adding a final concentration of about 8 mM N-acetylcysteine to the cell culture production medium.
20 . The method of claim 16 , wherein the antibody is selected from the group consisting of an anti-TNFα antibody, an anti-IL-18 antibody, and an anti-IL-12 antibody.