Methods of cell culture
AssigneeMomenta Pharmaceuticals, Inc.
InventorHolly Prentice
Life of this patent
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Abstract
Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described.
Claims35
A method of producing a recombinant protein preparation having a target level of one or more mature glycans, the method comprising:
- (a) providing a cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising putrescine under conditions in which the cell expresses the recombinant protein;
- (c) harvesting a preparation of the recombinant protein produced by the cell;
- and
- (d) measuring a level of one or more mature glycans in the preparation of the recombinant protein, wherein the preparation has the target level of the one or more mature glycans.
A method of producing a recombinant protein preparation having a target value of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans, the method comprising:
- (a) providing a cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising putrescine under conditions in which the cell expresses the recombinant protein;
- and
- (c) harvesting a preparation of the recombinant protein produced by the cell, wherein the preparation has the target value of the one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans.
A method of producing a recombinant protein preparation, the method comprising:
- (a) providing a target value of one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans;
- (b) providing a cell genetically engineered to express a recombinant protein;
- (c) culturing the cell in a culture medium comprising putrescine under conditions in which the cell expresses the recombinant protein;
- (d) harvesting a preparation of the recombinant protein produced by the cell;
- and
- (e) formulating the preparation into a drug product if the preparation meets the target value of the one or more of fucosylated glycans, galactosylated glycans, high mannose glycans, and sialylated glycans.
A method of increasing a level of one or more of fucosylated glycans, galactosylated glycans, and sialylated glycans in a recombinant protein preparation, the method comprising:
- (a) providing a cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising an elevated level of putrescine under conditions in which the cell expresses the recombinant protein;
- and
- (c) harvesting a preparation of the recombinant protein produced by the cell, wherein the preparation has an increased level of one or more of fucosylated glycans, galactosylated glycans, and sialylated glycans, relative to a corresponding level in a preparation of the recombinant protein produced using the medium not comprising the elevated level of putrescine.
A method of decreasing a level of one or more high mannose glycans in a recombinant protein preparation, the method comprising:
- (a) providing a cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising an elevated level of putrescine under conditions in which the cell expresses the recombinant protein;
- and
- (c) harvesting a preparation of the recombinant protein produced by the cell, wherein the preparation has a decreased level of high mannose glycans relative to a corresponding level in a preparation of the recombinant protein produced using the medium not comprising the elevated level of putrescine.
Dependencies are inferred from the claim text.