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US 9,217,168 B2Granted
Methods of cell culture
AssigneeMomenta Pharmaceuticals, Inc.
InventorHolly Prentice
Life of this patent
Estimating the term…
Abstract
Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using putrescine, are described.
Claims28
A method of producing a recombinant protein preparation having a target value of one or more of 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 0.1 mg/L to 10 mg/L putrescine under conditions in which the cell expresses the recombinant protein;
- and
- (c) harvesting a preparation of the recombinant protein produced by the cell that meets the target value of the one or more of galactosylated glycans, high mannose glycans, and sialylated glycans,
- wherein the target value of galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of galactosylated glycans, or sialylated glycans in a preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine;
- or
- wherein the target value of high mannose glycans is a level at least 10% lower than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine.
4 more elements
A method of producing a recombinant protein preparation, the method comprising:
- (a) providing a target value of one or more of 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 0.1 mg/L to 10 mg/L 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 galactosylated glycans, high mannose glycans, and sialylated glycans;
- wherein the target value of galactosylated glycans, or sialylated glycans is a level at least 10% higher than a level of galactosylated glycans or sialylated glycans in a preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine;
- or
- wherein the target value of high mannose glycans is a level at least 10% lower than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine.
6 more elements
A method of increasing a level of one or more of 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 0.1 mg/L to 10 mg/L 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 level of one or more of galactosylated glycans, and sialylated glycans that is at least 10% higher than a level of one or more of galactosylated glycans and sialylated glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine.
1 more element
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 0.1 mg/L to 10 mg/L 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 level of high mannose glycans that is at least 10% lower than a level of high mannose glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising 0.1 mg/L to 10 mg/L putrescine.
1 more element
Dependencies are inferred from the claim text.