Methods for increasing mannose content of recombinant proteins
AssigneeAmgen Inc.
InventorsChung-Jr HUANG, SR., 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.
Claims20
A cell culture media containing 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 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 cells during a growth phase with a cell culture media that does not contain mannose:
- initiating and maintaining a production phase in the cell culture by perfusion with a serum-free perfusion media containing mannose, wherein the mannose to total hexose ratio in the perfusion media is greater than 0 but less than 1.0.
The method according to 4, wherein the perfusion is accomplished by alternating tangential flow.
The method according to 11, wherein the perfusion is accomplished by alternating tangential flow using an ultrafilter or a microfilter.
Dependencies are inferred from the claim text.