Method of culturing a cell with a supplement

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 peptides having up to six amino acids, wherein at least one peptide comprises cysteine and at least one peptide comprises tyrosine. 
     
     
         2 . The cell culture medium of  claim 1 , wherein at least one of the remaining amino acids of the peptide are selected from the group consisting of alanine, glycine, serine, valine, proline, aspartic acid, and glutamic acid. 
     
     
         3 - 18 . (canceled) 
     
     
         19 . A method of culturing an animal cell, comprising contacting the cell with a basal cell culture medium under conditions supporting the cultivation of the cell and supplementing the basal cell culture medium with a feed comprising peptides having up to six amino acids, wherein at least one peptide comprises cysteine and at least one peptide comprises tyrosine. 
     
     
         20 . The method according to  claim 19 , wherein the basal cell culture medium is supplemented with the feed on more than one day. 
     
     
         21 . The method of  claim 20 , wherein the supplementation of the basal cell culture medium occurs either from day 0, day 1, day 2, day 3, day 4 or day 5 after starting the cell culture, and is supplemented daily thereafter through day 13 or 14. 
     
     
         22 . The method of  claim 20 , wherein the supplementation of the basal cell culture medium is at about 1-10%, or about 5-20%, of the total starting volume of the basal cell culture medium. 
     
     
         23 . (canceled) 
     
     
         24 . The method of  claim 20 , wherein the cell produces an immunoglobulin or a fragment thereof. 
     
     
         25 . The method of  claim 24 , wherein the cell produces greater than 3000 mg/L of immunoglobulin or a fragment thereof. 
     
     
         26 . A method of preparing a cell culture medium, comprising combining a basal medium with a supplement comprising peptides, and with a carbohydrate and at least one amino acid or salt thereof, wherein the supplement comprises at least one peptide comprising cysteine and at least one peptide comprising tyrosine. 
     
     
         27 . The method of  claim 26 , wherein at least one of the remaining amino acids of the peptide other than cysteine or tyrosine, are selected from alanine, glycine, serine, valine, proline, aspartic acid, arginine, glutamine or glutamic acid. 
     
     
         28 - 40 . (canceled) 
     
     
         41 . A method for analyzing a cell culture medium feed, or reconstituted medium, the method comprising determining the presence or absence of a peptide comprising cysteine and a peptide comprising tyrosine in the medium, feed or reconstituted medium, wherein the peptides comprise up to six amino acids. 
     
     
         42 . The method for analyzing a serum-free, protein-free, hydrolysate-free, cell culture medium, concentrated feed, or reconstituted medium of  claim 41  wherein the analyzing is performed by a method selected from the group comprising: mass spectrometry (LCMS); capillary electrophoresis and HPLC. 
     
     
         43 - 68 . (canceled)