Method for culturing Chinese hamster ovary cells to improve production of recombinant proteins
AssigneeGenentech, Inc.
InventorsJennie P. Mather, Mary C. Tsao
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Abstract
A method for culturing mammalian cells to improve production of recombinant proteins is provided. The culture media contain high levels of selected amino acids and may be supplemented with various growth factors and trace elements. In addition, the media are optionally serum-free. The media are preferably used for suspension culturing of Chinese hamster ovary cells at high densities.
Claims5
A method for culturing Chinese hamster ovary cells to improve production of recombinant proteins produced by said cells comprising culturing the cells in a culture medium containing the following components:
- asparagine at a concentration of from about 300 mg/l to about 750 mg/l, aspartic acid at a concentration of from about 300 mg/l to about 650 mg/l, glycine at a concentration of from about 300 mg/l to about 500 mg/l, isoleucine at a concentration of from about 300 mg/l to about 500 mg/l, leucine at a concentration of from about 400 mg/l to about 700 mg/l, lysine at a concentration of from about 1000 mg/l to about 2250 mg/l, methionine at a concentration of from about 80 mg/l to about 350 mg/l, serine at a concentration of from about 500 mg/l to about 750 mg/l, threonine at a concentration of from about 550 mg/l to about 950 mg/l, tryptophan at a concentration of from about 75 mg/l to about 230 mg/l, tyrosine at a concentration of from about 175 mg/l to about 400 mg/l, and valine at a concentration of from about 240 mg/l to about 640 mg/l.
A method for culturing Chinese hamster ovary cells to improve production of recombinant proteins produced by said cells comprising culturing the cells in the following culture medium consisting essentially of the following components at the indicated concentrations:
- ______________________________________ Component Concentration (mg/l) ______________________________________ Calcium chloride, anhydrous 116.61 Cupric sulfate, pentahydrate 0.0012 Ferric nitrate, nonahydrate 0.05 Ferrous sulfate, heptahydrate 0.417 Potassium chloride 759.0 Magnesium sulfate, anhydrous 48.835 Magnesium chloride, anhydrous 143.05 Sodium phosphate, monobasic, 62.5 monohydrate Sodium phosphate, dibasic, anhydrous 71.02 Zinc sulfate, heptahydrate 0.4315 Linoleic acid 0.294 Lipoic acid 0.735 Putrescine, dihydrochloride 0.5635 Sodium pyruvate 385.0 Alanine 31.15 Arginine, monohydrochloride 780.5 Asparagine, monohydrate 300-750 Aspartic acid 300-650 Cysteine, monohydrochloride, 122.92 monohydrate Cystine, dihydrochloride 31.285 Glutamic acid 51.45 Glutamine 1606.0 Glycine 300-500 Histidine, monohydrochloride, 94.36 monohydrate Isoleucine 300-500 Leucine 400-700 Lysine, monohydrochloride 1000-2250 Methionine 80-350 Phenylalanine 50.36 Proline 120.75 Serine 500-750 Threonine 500-950 Tryptophan 75-230 Tyrosine, disodium salt, dihydrate 175-400 Valine 250-640 Biotin 0.0256 D-Calcium pantothenate 3.68 Choline chloride 50.86 Cyanocobalamin 4.76 Folic acid 13.5-35 i-Inositol 66.60 Nicotinamide 2.1295 Pyridoxal, monohydrochloride 2.000 Pyridoxine, monohydrochloride 0.217 Riboflavin 0.3330 Thiamine, monohydrochloride 3.190 Glucose 4300.0 Sodium bicarbonate 1220.0 Sodium chloride (350 mOsm) 7360.0 Pluronic F-68 Prill 1000.0 Gentamycin sulfate 100.0. ______________________________________
Dependencies are inferred from the claim text.