Cell culture improvements

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.

1 - 102 . (canceled) 
     
     
         103 . A fed batch method of producing an anti-TNFα antibody comprising
 a) culturing Chinese Hamster Ovary (CHO) cells comprising a nucleic acid encoding the anti-TNFα antibody in a cell culture comprising a cell culture production medium; and 
 b) feeding the CHO cells by adding a hydrolysate enrichment solution and a basal enrichment solution to the cell culture during a time period, 
 wherein the basal enrichment solution comprises a basal medium, asparagine, and glucose, and 
 wherein the hydrolysate enrichment solution comprises at least two different non-animal-based hydrolysates, 
 such that the anti-TNFα antibody is produced. 
 
     
     
         104 . A fed batch method of producing an anti-TNFα antibody comprising
 a) culturing CHO cells comprising a nucleic acid encoding the anti-TNFα antibody in a cell culture comprising a cell culture production medium comprising at least 2.0 g/L of glucose, wherein the concentration of glucose is controlled by adding glucose to the cell culture production medium as required to maintain a concentration of at least 2.0 g/L of glucose; and 
 b) feeding the CHO cells by adding a hydrolysate enrichment solution and a basal enrichment solution to the cell culture during a time period, 
 wherein the basal enrichment solution comprises a basal medium, asparagine, and glucose, and 
 wherein the hydrolysate enrichment solution comprises at least two different non-animal-based hydrolysates, 
 such that the anti-TNFα antibody is produced. 
 
     
     
         105 - 174 . (canceled) 
     
     
         175 . A method for producing at least about 1.5 g/L of an antibody from a mammalian cell culture comprising
 a) culturing mammalian cells comprising a nucleic acid encoding the antibody in a cell culture production medium; and   b) adding a combination feed solution having a pH of about 6.7 to 7.2 to the cell culture production medium, wherein the combination feed solution comprises glucose; a basal cell medium; an amino acid other than glutamine;   and at least two different non-animal based hydrosylates, such that at least about 1.5 g/L of the antibody is produced.   
     
     
         176 - 233 . (canceled)