Cell Culture Media Containing Combinations of Proteins

Abstract

The present invention relates to cell culture media containing combinations of proteins, as well as methods of making the cell culture media, and methods of using the cell culture media to improve growth characteristics of cultured cells.

1 . A method for enhancing cell growth of a cell in culture comprising the addition of a supplement to said cell culture medium, wherein said supplement comprises; a mixture of recombinant albumin and a transferrin related protein, wherein said recombinant albumin is
 i) produced in a plant,   ii) has less than about 1 EU of endotoxin/mg of albumin; and   iii) less than about 2% aggregated albumin; and wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5,000, to about 1 to 0.5.   
     
     
         2 . A method for enhancing the productivity of a cell that has been adapted to serum free media comprising the addition of a supplement to said serum free media, wherein said supplement comprises; a mixture of recombinant albumin and a transferrin related protein; wherein said recombinant albumin is
 i) produced in a plant,   ii) has less than about 1 EU of endotoxin/mg of albumin; and   iii) less than about 2% aggregated albumin; and wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5,000, to about 1 to 0.5.   
     
     
         3 . A method for improving the yield of a recombinant product produced from cells in culture, comprising the addition of a supplement to said culture wherein said supplement comprises a mixture of recombinant albumin and a transferrin related protein;
 wherein said recombinant albumin is i) produced in a plant, ii) has less than about 1 EU of endotoxin/mg of albumin; and iii) less than about 2% aggregated albumin; and   wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5,000 to about 1 to 0.5.   
     
     
         4 . A method for improving the viability of cells in culture in a bioreactor comprising the addition of
 a supplement to said culture in said bioreactor wherein said supplement comprises;   a mixture of recombinant albumin and a transferrin related protein;   wherein said recombinant albumin is i) produced in a plant, ii) has less than about 1 EU of endotoxin/mg of albumin; and iii) less than about 2% aggregated albumin; and   wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5,000 to about 1 to 0.5.   
     
     
         5 . A method for improving the viability of cells grown in serum free media comprising the addition of a supplement to said serum free media wherein said supplement comprises
 a mixture of recombinant albumin and a transferrin related protein;   wherein said recombinant albumin is i) produced in a plant, ii) has less than about 1 EU of endotoxin/mg of albumin; and iii) less than about 2% aggregated albumin; and   wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5,000 to about 1 to 0.5.   wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5,000 to about 1 to 0.5.   
     
     
         6 . The method  claim 2  or  3 , wherein the viability of the cell in culture is increased. 
     
     
         7 . The method of any of  claims 1  to  5 , wherein said supplement further comprises insulin. 
     
     
         8 . The method of any of  claims 1  to  5 , wherein said supplement further comprises, selenium and ethanolamine. 
     
     
         9 . The method of any of  claims 1 - 5 , wherein said cells are tissue culture cells. 
     
     
         10 . The method of any of  claims 1 - 5 , wherein said cells are CHO cells. 
     
     
         11 . The method of any of  claims 1 - 5 , wherein said cells are hybridoma cells. 
     
     
         12 . The method of  claim 3 , wherein said protein is an antibody. 
     
     
         13 . The method of any of  claims 1 - 5 , wherein said recombinant albumin comprises at least about 0.01% wt/wt of a heat shock protein. 
     
     
         14 . The method of  claim 13 , wherein said heat shock protein is a rice heat shock protein. 
     
     
         15 . The method of  claim 13 , wherein said heat shock protein is selected from said group consisting of Rice HSP70 genes, and rice endosperm lumenal binding protein. 
     
     
         16 . The method of  claim 13 , wherein said heat shock protein is selected from the group consisting of Rice (gb|ACJ54890.1|), EEC69073/OsI — 37938, and AAB63469. 
     
     
         17 . The method of  claim 13 , wherein said transferrin related protein is lactoferrin. 
     
     
         18 . The method of  claim 13 , wherein said transferrin related protein is transferrin. 
     
     
         19 . The method of  claim 18 , wherein said ratio of said transferrin to said recombinant albumin is about 1 to 50, to about 1 to 5000. 
     
     
         20 . The method of  claim 17 , wherein said lactoferrin is recombinant lactoferrin. 
     
     
         21 . The method of  claim 20 , wherein said ratio of said lactoferrin to said recombinant albumin is about 1 to 3, to about 1 to 0.33. 
     
     
         22 . The method of any of  claims 4  to  6  wherein said improvement in cell viability is greater than 30% compared to the summed effect of said recombinant albumin and said transferrin related protein when added alone when measured under the same culture conditions. 
     
     
         23 . A supplement comprising; a mixture of recombinant albumin and a transferrin related protein; wherein said recombinant albumin is i) produced in a plant, ii) has less than about 1 EU of endotoxin/mg of albumin; and iii) less than about 2% aggregated albumin; and wherein said supplement comprises a ratio (wt/wt) of said transferrin related protein to said recombinant albumin of about 1 to 5000 to about 1 to 0.5. 
     
     
         24 . The supplement of  claim 23 , wherein said recombinant albumin comprises at least about 0.01% wt/wt of a heat shock protein. 
     
     
         25 . The supplement of  claim 24 , wherein said heat shock protein is a rice heat shock protein. 
     
     
         26 . The supplement of  claim 24 , wherein said heat shock protein is selected from said group consisting of Rice HSP70 genes, and rice endosperm lumenal binding protein. 
     
     
         27 . The supplement of  claim 24 , wherein said heat shock protein is selected from the group consisting of Rice (gb|ACJ54890.1|), EEC69073/OsI — 37938, and AAB63469. 
     
     
         28 . The supplement of  claim 24 , wherein said supplement comprises at least about 0.04% wt/wt HSP70. 
     
     
         29 . The supplement of  claim 24 , wherein said transferrin related protein is lactoferrin. 
     
     
         30 . The supplement of  claim 24 , wherein said transferrin related protein is transferrin. 
     
     
         31 . The supplement of  claim 30 , wherein said ratio of said transferrin to said recombinant albumin is about 1 to 50, to about 1 to 5000. 
     
     
         32 . The supplement of  claim 31 , wherein said ratio of said lactoferrin to said recombinant albumin is about 1 to 3, to about 1 to 0.33.