Methods for increasing mannose content of recombinant proteins
AssigneeAmgen Inc.
InventorsJian Wu, Sean Davern, Simina Crina Petrovan, Michael Charles Brandenstein, Katherine Rose Lindahl, Shawn Erik Lillie
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Abstract
The present invention relates to methods of modulating the mannose content of recombinant proteins.
Claims26
A method of modulating the amount of the mannose-5 glycoform of an Immunoglobulin G2 (IgG2) molecule in an IgG2 composition, wherein said IgG2 composition is produced by a Chinese Hamster Ovary (CHO) cell culture, comprising:
- (a) establishing a cell culture of CHO cells in a bioreactor, wherein the CHO cell produces an IgG2 molecule;
- (b) limiting the amount of glucose in the bioreactor, wherein the concentration of the glucose in the spent medium of the bioreactor is from about 0 to 3 g/L;
- (c) supplementing the cell culture with a feeding medium comprising galactose, such that the concentration of galactose in the resulting spent medium of the bioreactor is above 2.5 g/L;
- and
- (d) harvesting and purifying the IgG2 composition from the cell culture.
A method of producing an immunoglobulin G2 (IgG2) composition by a Chinese Hamster Ovary (CHO) cell culture, comprising:
- (a) establishing a cell culture of CHO cells in a bioreactor, wherein the CHO cell produces an IgG2 molecule;
- (b) limiting the amount of glucose in the bioreactor, wherein the concentration of the glucose in the spent medium of the bioreactor is from about 0 to 3 g/L;
- (c) supplementing the cell culture with a feeding medium comprising galactose, such that the concentration of galactose in the resulting spent medium of the bioreactor is above 2.5 g/L;
- and
- (d) harvesting and purifying the IgG2 composition from the cell culture.
A method of producing an immunoglobulin G2 (IgG2) composition by a Chinese Hamster Ovary (CHO) cell culture, wherein said IgG2 is denosumab, comprising:
- (a) establishing a cell culture of CHO cells in a bioreactor, wherein the CHO cell produces denosumab molecule;
- (b) limiting the amount of glucose in the bioreactor, wherein the concentration of the glucose in the spent medium of the bioreactor is from about 0 to 3 g/L;
- (c) supplementing the cell culture with a feeding medium comprising galactose, such that the concentration of galactose in the resulting spent medium of the bioreactor is above 2.5 g/L;
- and
- (d) harvesting and purifying the denosumab composition from the cell culture.
Dependencies are inferred from the claim text.