Cell Culture Media for Enhanced Protein Production

Abstract

Culture media for in vitro cell culture which contain substantially saturated amounts of selected amino acids improve protein production, constrain cell growth and extend cell longevity, methods for the production and use of such media, and systems fox the production of protein utilizing such media and methods.

1 - 32 . (canceled) 
   
   
       33 . A method for culturing animal cells, comprising:
 (a) providing an animal cell;   (b) providing a cell culture medium comprising:
 1) a total of from approximately 5.5 to 20 grams per liter of a plurality of amino acids; and 
 2) at least one carbohydrate; and 
   (c) growing the animal cell of (a) in the medium of (b), wherein the utilization rate of the carbohydrate source as an energy source by the animal cell is altered as compared to the utilization rate in a conventional medium, thereby culturing cells.   
   
   
       34 . The method of  claim 33 , wherein the animal cell utilizes an alternative source of carbon. 
   
   
       35 . The method of  claim 33 , wherein the utilization rate of the carbohydrate source as an energy source by the animal cell is lower as compared to the utilization rate in a conventional medium. 
   
   
       36 . The method of  claim 33 , wherein the animal cell does not utilize the carbohydrate source as an energy source. 
   
   
       37 . The method of  claim 33 , wherein the cell produces at least one protein of interest and production of the at least one protein of interest is increased relative to the production of the at least one protein of interest produced by the same cell grown in the conventional medium. 
   
   
       38 . The method of  claim 35 , wherein the conventional medium is selected from the group consisting of: DMEM, DMEM/F12, eRDF, and RPMI. 
   
   
       39 . The method of  claim 33 , wherein the cell produces at least one protein of interest and production of the at least one protein of interest is increased at least about two-fold relative to the production of the at least one protein of interest produced by the same cell grown in the conventional medium. 
   
   
       40 . The method of  claim 33 , wherein the cell produces at least one protein of interest and production of the at least one protein of interest is increased at least about five-fold relative to the production of the at least one protein of interest produced by the same cell grown in the conventional medium. 
   
   
       41 . The method of  claim 33 , wherein the cell produces at least one protein of interest and production of the at least one protein of interest is increased at least about ten-fold relative to the production of the at least one protein of interest produced by the same cell grown in the conventional medium. 
   
   
       42 . The method of  claim 33 , wherein the cell remains viable for a longer period of time in continuous culture without the replenishment or exchange of growth medium as compared to the time the same type of cell remains viable when grown in the conventional medium. 
   
   
       43 . The method of  claim 42 , wherein the conventional medium is selected from the group consisting of DMEM, DMEM/F12, eRDF, and RPMI. 
   
   
       44 . The method of  claim 33 , wherein the cell remains viable for at least twice as long as the same type of cell grown in the conventional medium. 
   
   
       45 . The method of  claim 33 , wherein the cell remains viable for at least five times as long as the same type of cell grown in the conventional medium. 
   
   
       46 . The method of  claim 33 , wherein the cell is a hybridoma cell or a myeloma cell and the at least one protein of interest is an antibody. 
   
   
       47 . The method of  claim 33 , wherein the cell comprises a recombinant protein expression vector and the at least one protein of interest is a recombinant protein expressed by the vector in the cell. 
   
   
       48 . The method of  claim 47 , wherein the cell is a mammalian cell. 
   
   
       49 . The method of  claim 48 , wherein the mammalian cell is selected from the group consisting of a CHO cell, a BHK cell, a COS cell, and a Namalwa cell.