FEED SUPPLEMENT FOR MAMMALIAN CELL CULTURE AND METHODS OF USE
An improved feed supplement for culture of mammalian cells used to produce proteins is provided. The improved supplement is devoid of animal-derived components and protein hydrolysates. The invention also provides methods of using the supplement in production of a therapeutic proteins, such as an antibody. In some embodiments, the antibody is an anti-human IL-23p19 antibody.
1 . A cell feed supplement concentrate comprising components 1-47 of Table 3 and;
a) sodium selenite; or
b) vitamin E.
2 . The supplement of claim 1 comprising sodium selenite and vitamin E.
3 . The supplement of claim 1 , wherein the sodium selenite is present at approximately 0.3 mg/L.
4 . The supplement of claim 1 , wherein the vitamin E is present at approximately 30.2 mg/L.
5 . A method of producing a protein comprising culturing cells in growth medium supplemented with the supplement of claim 1 .
6 . A method of producing a protein comprising:
a) growing mammalian cells expressing the protein in culture;
b) supplementing the culture with the supplement of claim 1 ; and
c) recovery of the protein from the culture.
7 . The method of claim 6 further comprising shifting the temperature of the culture from 37° C. to 34° C.
8 . The method of claim 7 wherein the shifting of the temperature is performed on day 3, day 4 or day 5 after inoculation.
9 . The method of claim 6 wherein the supplementing step (b) is repeated an additional two or more times.
10 . The method of claim 10 wherein the supplementing steps are performed on days 3, 5 and 10 after inoculation.
11 . The method of claim 5 or 6 wherein the cells are CHO cells.
12 . The method of claim 5 or 6 wherein the protein is an antibody or antigen-binding fragment thereof.
13 . The method of claim 12 wherein the antibody or antigen-binding fragment thereof comprises a human IgG constant domain.
14 . The method of claim 12 wherein the antibody, or antigen-binding fragment thereof, specifically binds to human IL-23p19.
15 . The method of claim 14 wherein the antibody, or antigen-binding fragment thereof, comprises at least one heavy chain CDR, and at least one light chain CDR, of antibody hu13B8b disclosed in International Pat. Appl. Pub. No. WO 2008/103432.
16 . A method of producing a protein comprising:
a) growing mammalian cells expressing the protein in culture;
b) supplementing the culture with sodium selenite or vitamin E; and
c) obtaining the protein from the cell culture.
17 . The method of claim 16 wherein the culture is supplemented with sodium selenite and vitamin E.
18 . The method of claim 16 , wherein the sodium selenite is added to the culture to give a final concentration of approximately 0.02 mg/L.
19 . The method of claim 16 , wherein the vitamin E is added to the culture to give a final concentration of approximately 2 mg/L.
20 . The method of claim 16 , wherein the sodium selenite is added to the culture to give a final concentration of approximately 0.02 mg/L and the vitamin E is added to the culture to give a final concentration of approximately 2 mg/L.