Fed-batch method of making human anti-TNF-alpha antibody

1. A fed-batch method of making a human anti-TNFα antibody comprising a light chain variable region (LCVR) comprising the sequence of SEQ ID NO:1 and a heavy chain variable region (HCVR) comprising the sequence of SEQ ID NO:2, said method comprising culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding said LCVR and HCVR of said anti-TNFα antibody in a cell culture production medium in large-scale,

wherein glucose concentration in said cell production medium is monitored, the glucose concentration in said medium decreases to below 2 g/L, and glucose is added to said medium when the glucose concentration in said medium decreases to below 2 g/L, or the glucose concentration in said medium is monitored and glucose is added to said medium to maintain the glucose concentration in said medium at a concentration of at least 2 g/L but no greater than 7 g/L, such that said anti-TNFα antibody is produced, and wherein said produced antibody is further affinity purified using a Protein A resin.

2. The method of claim 1 , wherein said produced antibody has a titer of at least 1.5 g/L in said cell culture production medium.

3. The method of claim 2 , wherein said culturing lasts 9-15 days.

4. The method of claim 1 , wherein said produced antibody has a titer of at least 2 g/L in said cell culture production medium.

5. The method of claim 1 , wherein said cell culture production medium is a serum-free medium.

6. The method of claim 5 , wherein said serum-free medium comprises at least one non-animal-based hydrolysate selected from the group consisting of a yeast-based hydrolysate, a soy-based hydrolysate, and a combination thereof.

7. The method of claim 5 , wherein said serum-free medium comprises galactose and manganese.

8. The method of claim 1 , wherein said CHO cells are cultured at a temperature ranging from 32° C. to 38° C.

9. The method of claim 8 , wherein said temperature is maintained as a constant temperature throughout said production method, said temperature being selected from a temperature range of 32° C. to 38° C.

10. A fed-batch production method of making adalimumab, comprising culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding adalimumab in a cell culture production medium in large-scale, wherein glucose concentration in said medium is monitored, the glucose concentration in said medium decreases to below 2 g/L, and glucose is added to said medium when the glucose concentration in said medium decreases to below 2 g/L, or the glucose concentration in said medium is monitored and glucose is added to said medium to maintain the glucose concentration in said medium at a concentration of at least 2 g/L but no greater than 7 g/L, such that adalimumab is produced, and wherein said produced adalimumab is further affinity purified using a Protein A resin.

11. The method of claim 10 , wherein said produced antibody has a titer of at least 1.5 g/L in said cell culture production medium.

12. The method of claim 11 , wherein said culturing lasts 9-15 days.

13. The method of claim 11 , wherein said cell culture production medium is a serum-free medium.

14. The method of claim 13 , wherein said serum-free medium comprises at least one non-animal-based hydrolysate selected from the group consisting of a yeast-based hydrolysate, a soy-based hydrolysate, and a combination thereof.

15. The method of claim 13 , wherein said serum-free medium comprises galactose and manganese.

16. The method of claim 10 , wherein said produced antibody has a titer of at least 2 g/L in said cell culture production medium.

17. The method of claim 16 , wherein said culturing lasts 9-15 days.

18. The method of claim 16 , wherein said cell culture production medium is a serum-free medium.

19. The method of claim 18 , wherein said serum-free medium comprises at least one non-animal-based hydrolysate selected from the group consisting of a yeast-based hydrolysate, a soy-based hydrolysate, and a combination thereof.

20. The method of claim 18 , wherein said serum-free medium comprises galactose and manganese.

21. The method of claim 11 , wherein said produced antibody has a titer of at least 2.5 g/L in said cell culture production medium.

22. The method of claim 21 , wherein said culturing lasts 9-15 days, and wherein (a) said cell culture production medium is a serum-free medium and comprises at least one non-animal-based hydrolysate, or (b) said cell culture production medium is a serum-free medium comprising galactose and manganese.

23. The method of claim 10 , wherein said produced antibody has a titer of at least 4 g/L in said cell culture production medium and (a) said cell culture production medium is a serum-free medium and comprises at least one non-animal-based hydrolysate, or (b) said cell culture production medium is a serum-free medium comprising galactose and manganese.

24. The method of claim 10 , wherein said CHO cells are cultured at a temperature ranging from 32° C. to 38° C.

25. A fed-batch method of producing adalimumab, comprising expressing adalimumab in Chinese Hamster Ovary (CHO) cells in a cell culture production medium in large -scale, wherein said CHO cells comprise an expression vector encoding adalimumab, wherein glucose concentration in said medium is monitored, and the glucose concentration in said medium drops to a level between 1 and 5 g/L, and glucose is added to said medium to maintain the glucose concentration in said medium at least 1-5g/L, and wherein adalimumab is further affinity purified using a Protein A resin.

26. The method of claim 25 , wherein said produced antibody has a titer of at least 1.5 g/L in said cell culture production medium.

27. The method of claim 25 , wherein said produced antibody has a titer of at least 2 g/L in said cell culture production medium.

28. The method of claim 25 , wherein said produced antibody has a titer of at least 2.5 g/L in said cell culture production medium.