Methods for increasing mannose content of recombinant proteins
AssigneeAmgen Inc.
InventorsChung-Jr Huang, Xiaoming Yang
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Abstract
The present invention relates to methods of upregulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture by manipulating the mannose to total hexose ratio in the cell culture media formulation.
Claims19
A cell culture media comprising mannose, wherein the mannose to total hexose ratio in the cell culture media is greater than 0 but less than 1.0.
A method for increasing the high mannose glycoform content of a recombinant protein during a mammalian cell culture process comprising:
- establishing a mammalian cell culture in a bioreactor with a cell culture media that does not contain mannose;
- and
- maintaining the mammalian cell culture with a cell culture media comprising
- mannose, wherein the mannose to total hexose ratio in the cell culture media is greater than 0 but less than 1.0, wherein the high mannose glycoform content of the recombinant protein is increased compared to a mammalian cell culture wherein the mammalian cell culture is not subjected to a cell culture media containing a mannose to total hexose ratio of greater than 0 and less than 1.0.
A method for upregulating the high mannose glycoform content of a recombinant protein during a mammalian cell culture process comprising:
- establishing a mammalian cell culture in a bioreactor with a cell culture media that does not contain mannose;
- growing the mammalian cell culture in a growth phase with a cell culture media that does not contain mannose;
- initiating and maintaining a production phase in the mammalian cell culture by perfusion with a cell culture media comprising mannose, wherein the mannose to total hexose ratio in the cell culture media is greater than 0 but less than 1.0, wherein the high mannose glycoform content of the recombinant protein is increased compared to a mammalian cell culture wherein the mammalian cell culture is not subjected to a cell culture media comprising mannose with a mannose to total hexose ratio greater than 0 and less than 1.0.
The method of 11, wherein the alternating tangential flow comprises an ultrafilter or a microfilter.
Dependencies are inferred from the claim text.