Methods for modifying glycosylation using manganese
AssigneeAres Trading S.A.
InventorsThomas Solacroup, Matthieu Stettler, Martin Jordan, Hervé Broly
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Abstract
The present invention relates to a cell culture method for adjusting the proportion of G0, G1, G2, and high-mannose galactosylation variants in a population of a recombinant protein produced by a culture of cells in a cell culture medium by supplementing the cell culture medium with manganese at one or more days of the cell culture method duration.
Claims15
A cell culture method for adjusting the proportion of G0, G1, G2, and high-mannose glycosylation variants in a population of a recombinant protein produced by a culture of cells in a cell culture medium, the cell culture method comprising:
- i) initiating the cell culture method in a manganese-free, galactose-free cell culture medium;
- and
- ii) supplementing the cell culture medium with manganese (Mn) at one or more days of the cell culture method duration and wherein the cell culture medium remains galactose free for the cell culture method duration, thereby adjusting the proportion of G0, G1, G2, and high-mannose glycosylation variants in the population of the recombinant protein in comparison to a population of the recombinant protein produced in cell culture that is not supplemented with manganese and/or contains galactose.
A method of making a recombinant protein in a cell culture, comprising:
- i) initiating the cell culture method in a manganese-free, galactose-free cell culture medium;
- ii) growing a host cell in a cell culture medium,
- iii) expressing the recombinant protein in the host cell,
- iv) supplementing the cell culture medium with manganese at one or more days of the cell culture method duration and wherein the cell culture medium remains galactose free for the cell culture method duration, thereby adjusting the proportion of G0, G1, G2, and high-mannose glycosylation variants in the population of the recombinant protein in comparison to a population of the recombinant protein produced in cell culture that is not supplemented with manganese and/or contains galactose, and
- v) purifying the recombinant protein from the cell culture.
Dependencies are inferred from the claim text.