Methods for increasing mannose content of recombinant proteins

摘要

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.

主权项

1. A method for increasing a high mannose glycoform content of a recombinant protein during a cell culture process comprising:
establishing a cell culture comprising mammalian cells in a bioreactor with a cell culture media that does not contain mannose; growing said mammalian cells in a growth phrase with a cell culture media that does not contain mannose; initiating and maintaining a production phase in said cell culture by perfusion with a serum-free perfusion media comprising mannose to produce said recombinant protein, wherein a mannose to total hexose ratio in said serum-free perfusion media is greater than 0 but less than 1.0, wherein said high mannose glycoform content of said recombinant protein is increased compared to a cell culture wherein said cell culture is not subjected to a serum-free perfusion media comprising mannose with a mannose to total hexose ratio greater than 0 and less than 1.0.