Cell culture media for enhanced protein production

Abstract

A cell culture medium is provided which constrains cell growth and enhances antibody production. The high glucose medium of the invention is preferably saturated at 40° C. with essential amino acids.

What we claim:  
     
         1 . In a cell culture medium comprising an aqueous solution of amino acids and a carbohydrate energy source for cells cultured in said medium, 
 the improvement wherein said aqueous solution 
 (i) is substantially saturated with said amino acids at a temperature of 30° to 50° C.;  
 (ii) has an osmolarity of 320 to 450 mOsm;  
 (iii) contains 5.50 to 20 grams per liter of amino acids and 5.50 to 20 grams per liter of said carbohydrate energy source  
 and wherein the dry weight of the amino acids in solution in said medium comprises at least 20% of the total dry weight of all solid components present in said medium.  
   
     
     
         2 . The  claim 1  cell culture medium in which said carbohydrate energy source is glucose.  
     
     
         3 . The  claim 1  cell culture medium in which said osmolarity of from 320 to 450 mOsm is provided at least in part by sodium salt.  
     
     
         4 . The  claim 1  or  claim 2  cell culture medium in which said osmolarity of from 320 to 450 mOsm is provided at least in part by a sodium chloride osmolyte.  
     
     
         5 . The  claim 1  cell culture medium further comprising a suspension of undissolved amino acids in said aqueous solution substantially saturated with amino acids 
 wherein said substantial saturation is maintained as cell growth consumes amino acids in solution in said aqueous medium.  
 
     
     
         6 . In an aqueous basal cell culture medium, the improvement which comprises providing in said basal medium 
 (i) a concentration of one or more amino acids reactive with cell transport system A to provide a total amino acid concentration of 5.50 to 20 grams per liter;    (ii) an osmolarity of from 320 to 420 mOsm     said osmolarity being provided by a sodium salt osmolyte.    
     
     
         7 . The  claim 6  cell culture medium in which the osmolyte (ii) is sodium chloride.  
     
     
         8 . The  claim 6  cell culture medium in which said one or more amino acids reactive with cell transport system A are zwitterionic amino acids.  
     
     
         9 . The  claim 6  cell culture medium in which said one or more amino acids reactive with cell transport system A are selected from the group consisting of alanine, glycine, histidine, methionine, proline and serine.  
     
     
         10 . The  claim 6  or  claim 7  or  claim 8  cell culture medium in which said basal cell culture medium is RPMI, DMEM, Ham's F12, RDF or eRDF.  
     
     
         11 . The cell culture medium 28101, 28102 or 28103.  
     
     
         12 . The method which comprises culturing a cell in the culture medium of  claim 1 ,  claim 2 , or  claim 3 , or  claim 11 .  
     
     
         13 . A method which comprises: 
 (i) providing the cell culture medium of  claim 1  or  claim 2  or  claim 3  or claim  11 ;    (ii) culturing a CHO cell containing a recombinant expression vector in the medium provided in step (i); and    (iii) recovering the recombinant protein expressed by said CHO cell.    
     
     
         14 . The  claim 13  method in which the CHO cell containing a recombinant expression vector is a dicistronic DHFR intron expression vector.  
     
     
         15 . A method which comprises culturing a CHO cell in culture medium 28103.  
     
     
         16 . The method of  claim 13  wherein said cell cultured in said culture medium is a hybridoma.  
     
     
         17 . The method which comprises culturing a mammalian cell in the culture medium of  claim 1 ,  claim 2  or  claim 3  or  claim 11 .  
     
     
         18 . The method which comprises culturing a mammalian cell having an expression vector for a recombinant protein in a culture medium of  claim 1 ,  claim 2  or  claim 3  or  claim 11 .  
     
     
         19 . The method which comprises culturing a CHO cell or a BHK cell or a CO5 cell or a Namaliva cell having expression vector for a recombinant protein in a culture medium of  claim 1 ,  claim 2 ,  claim 3  or  claim 11 .  
     
     
         20 . The method which comprises culturing a cell in the culture medium of  claim 6  or  claim 7  or  claim 8  or  claim 9 .  
     
     
         21 . The method which comprises culturing a mammalian cell in the culture medium of  claim 7  or  claim 8  or  claim 9 .  
     
     
         22 . The method which comprises culturing a mammalian cell having an expression vector for a recombinant protein in a culture medium of  claim 7  or  claim 8  or  claim 9 .  
     
     
         23 . The method which comprises culturing a CHO cell or a BHK cell or a CO5 cell or a Namaliva cell having expression vector for a recombinant protein in a culture medium of  claim 7  or  claim 8  or  claim 9 .  
     
     
         24 . A dry mixture of cell culture medium components comprising amino acids, carbohydrates and vitamins, 
 wherein said amino acids component comprise at least 20% of the dry weight of said mixture.    
     
     
         25 . The  claim 24  mixture wherein said amino acids component comprises from 30% to 50% of the dry weight of said mixture.