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US 8,956,830 B2Granted
Methods of cell culture
AssigneeMomenta Pharmaceuticals, Inc.
InventorsHolly Prentice, Rasheed Tijani, Brett Belongia
Life of this patent
Estimating the term…
Abstract
Polypeptide preparations having target levels of glycans, and methods of producing such polypeptide preparations using DMSO, are described.
Claims57
A method of producing a recombinant protein preparation having target values of high mannose glycans and fucosylated glycans, the method comprising:
- (a) providing a mammalian cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising dimethylsulfoxide (DMSO) 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 values of the high mannose glycans and fucosylated glycans, wherein the target value of high mannose glycans is a level at least 20% higher than a level of high mannose glycans and the target value of fucosylated glycans is a level at least 10% lower than in a preparation produced by culturing the cell in the medium not comprising DMSO.
1 more element
A method of producing a recombinant protein preparation, the method comprising:
- (a) providing target values of mannose glycans and fucosylated glycans;
- (b) providing a mammalian cell genetically engineered to express a recombinant protein;
- (c) culturing the cell in a culture medium comprising DMSO 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 values of the high mannose glycans and fucosylated glycans, and wherein the target value of high mannose glycans is a level at least 20% higher than a level of high mannose glycans and the target value of fucosylated glycans is a level at least 10% lower than in a preparation produced by culturing the cell in the medium not comprising DMSO.
3 more elements
A method of increasing a level of one or more of sialylated glycans and 3,3,1,0,0 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 DMSO 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 sialylated glycans and 3,3,1,0,0 glycans relative to a level of one or more of sialylated glycans and 3,3,1,0,0 glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising DMSO.
1 more element
A method of decreasing a level of one or more fucosylated 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 DMSO 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 at least 10% of one or more fucosylated glycans relative to a level of one or more fucosylated glycans in a preparation of the recombinant protein produced by culturing the cell in the medium not comprising DMSO.
1 more element
A method of producing a recombinant protein preparation having a target value of high mannose glycans, the method comprising:
- (a) providing a mammalian cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising DMSO 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 high mannose glycans, wherein the target value is a level at least 20% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO.
1 more element
A method of producing a recombinant protein preparation, the method comprising:
- (a) providing a mammalian cell genetically engineered to express a recombinant protein;
- (b) culturing the cell in a culture medium comprising DMSO under conditions in which the cell expresses the recombinant protein;
- (c) harvesting a preparation of the recombinant protein produced by the cell;
- and
- (d) formulating the preparation into a drug product if the preparation comprises a level of high mannose glycans at least 20% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO.
2 more elements
A method of producing a recombinant therapeutic protein preparation, the method comprising:
- (a) providing a mammalian cell genetically engineered to express a recombinant therapeutic protein;
- (b) culturing the cell in a culture medium comprising 1-5% DMSO under conditions in which the cell expresses the recombinant therapeutic protein;
- (c) harvesting a preparation of the recombinant protein produced by the cell;
- and
- (d) formulating the preparation into a therapeutic drug product if the preparation comprises a level of high mannose glycans at least 20% higher than a level of high mannose glycans in a preparation produced by culturing the cell in the medium not comprising DMSO.
2 more elements
Dependencies are inferred from the claim text.