Cell culture medium comprising small peptides

Abstract

Cell culture media, concentrated media and feeds, methods of manufacturing cell culture media and feeds, and methods of culturing cells are provided. One or more small peptides, including dipeptides are added to the cell culture media to provide improved stability and improved conditions for culturing cells.

1 . A cell culture medium comprising at least one small peptide, said peptide comprising at least two amino acids, wherein at least one of the amino acids is a cysteine or a tyrosine. 
     
     
         2 . The cell culture medium of  claim 1 , wherein at least one of the remaining amino acids of the small peptide are selected from the group consisting of alanine, glycine, serine, valine, proline, aspartic acid, and glutamic acid. 
     
     
         3 . The cell culture medium of  claim 2 , wherein at least one of the remaining amino acids of the small peptide is alanine or glycine. 
     
     
         4 . The cell culture medium of  claim 1 , wherein at least one of the small peptides is a dipeptide selected from the group consisting of X-tyrosine, X-cysteine, tyrosine-X, and cysteine-X, or a salt thereof, and wherein X is selected from the group consisting of alanine, glycine, serine, valine, proline, aspartic acid, and glutamic acid, and/or, and wherein the N-terminal amino acid of the one or small peptides has a free amino group. 
     
     
         5 . The cell culture medium of  claim 1 , wherein X is alanine or glycine. 
     
     
         6 . The cell culture medium of  claim 1 , wherein the cell culture medium is a liquid solution. 
     
     
         7 . The cell culture medium of  claim 1 , wherein the cell culture medium is a dry powder or a granulated dry powder. 
     
     
         8 . The cell culture medium of  claim 1 , further comprising a carbohydrate and an amino acid, or a salt thereof. 
     
     
         9 . The cell culture medium of  claim 8 , wherein the carbohydrate is a hexose. 
     
     
         10 . The cell culture medium of  claim 8 , wherein the amino acid, or salt thereof, is one or more of arginine, asparagine, aspartic acid, cysteine, glutamic acid, histidine, isoleucine, lysine, methionine, phenylalanine, proline, hydroxyproline, serine, threonine, tryptophan, tyrosine, and valine. 
     
     
         11 . The cell culture medium of  claim 1 , further comprising a vitamin, a salt, a buffering agent, or an inorganic element. 
     
     
         12 . The cell culture medium of  claim 1 , wherein the cell culture medium does not contain a lipid, a hydrolysate or a fraction thereof, or a growth factor. 
     
     
         13 . The cell culture medium of  claim 4 , wherein the cell culture medium does not contain a protein. 
     
     
         14 . The cell culture medium of  claim 4 , wherein the cell culture medium is concentrated as a 2× or greater formulation. 
     
     
         15 . The cell culture medium of  claim 4 , wherein the one or more dipeptides are alanyl tyrosine, and/or alanyl cysteine, and/or alanyl cystine dimer. 
     
     
         16 . The cell culture medium of  claim 4 , wherein the one or more dipeptides are present at a concentration of about 1 g/L to about 16 g/L. 
     
     
         17 . The cell culture medium of  claim 4 , wherein the one or more dipeptides are present at a concentration of about 2.5 g/L to about 8.5 g/L. 
     
     
         18 . The cell culture medium of  claim 6 , wherein the liquid solution stored at 2-8° C. remains free of precipitate for over 12 months. 
     
     
         19 - 49 . (canceled) 
     
     
         50 . The cell culture medium of  claim 9 , wherein the hexose is selected from the group consisting of glucose, galactose, fructose and maltose. 
     
     
         51 . A cell culture medium, concentrated feed, or reconstituted medium comprising a small peptide comprising cysteine or tyrosine, wherein said cell culture medium, concentrated feed, or reconstituted medium is serum-free, protein-free and/or hydrolysate-free. 
     
     
         52 . The cell culture medium, concentrated feed, or reconstituted medium of  claim 51  wherein said small peptide comprises two, three, four, five or six amino acids. 
     
     
         53 . The cell culture medium, concentrated feed, or reconstituted medium of  claim 1  that supports the production of a recombinant protein. 
     
     
         54 . The cell culture medium, concentrated feed, or reconstituted medium of  claim 53  wherein the recombinant protein is an immunoglobulin or a fragment thereof. 
     
     
         55 . The composition comprising the cell culture medium, concentrated feed, or reconstituted medium of  claim 1  and a cell. 
     
     
         56 - 62 . (canceled) 
     
     
         63 . Use of the cell culture medium, concentrated feed, or reconstituted medium of  claim 1  to produce a virus or a recombinant protein. 
     
     
         64 . (canceled) 
     
     
         65 . The cell culture medium of  claim 1 , wherein the small peptide is either a dipeptide, a tripeptide, a tetrapeptide, a pentapeptide, a hexapeptide, a heptapeptide, an octapeptide, a nonapeptide, or a decapeptide. 
     
     
         66 . The cell culture medium of  claim 1 , wherein the small peptide is at least about two to ten amino acids in length. 
     
     
         67 . The cell culture medium of  claim 1 , wherein the small peptide is at least about ten amino acids in length. 
     
     
         68 . (canceled)