Control of protein glycosylation by culture medium supplementation and cell culture process parameters
Inventors
Yang, William • Huang, Yao-Ming • MCELEARNEY, Kyle • Tescione, Lia • Lambropoulos, James • Zhang, An • Tsang, Valerie • Ryll, Thomas • Lee, Sangil • Lee, Dae Sung
Assignees
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Publication Number
US-9944968-B2
Publication Date
2018-04-17
Expiration Date
Abstract
The present invention pertains to a cell culture medium comprising media supplements that are shown to control recombinant protein glycosylation and/or cell culture in a controlled or modulated (shifted) temperature to control recombinant protein glycosylation and/or cell culture with controlled or modulated seed density to control recombinant protein glycosylation, and methods of using thereof. The present invention further pertains to a method of controlling or manipulating glycosylation of a recombinant protein of interest in a large scale cell culture.
Core Innovation
The invention describes a method of altering the glycosylation pattern of a recombinant glycoprotein produced in cell culture by culturing eukaryotic cells engineered to express a recombinant glycoprotein of interest in a cell culture medium. The cell culture medium is supplemented with an additive comprising mycophenolic acid acyl glucuronide, and the glycosylation pattern of the recombinant glycoprotein is altered relative to the same recombinant glycoprotein produced by the same cells in the same cell culture medium without the additive.
The disclosed approach is directed to changing glycosylation attributes such as afucosylation, galactosylation, α-gal, and N-glycolylneuraminic acid (NGNA). The alteration can include reduced FcγRIIIa binding and reduced antibody-dependent cell-mediated cytotoxicity (ADCC) associated with the modified glycan profile.
The partial content also describes controlling recombinant glycoprotein glycosylation through cell culture medium supplementation and modulation of culture conditions. In addition to mycophenolic acid acyl glucuronide, described additives include mycophenolic acid, insulin, copper(II) sulfate, hypoxanthine/guanine, glucosamine, and galactose and mixtures, together with modulating culture conditions such as temperature and seed density.
Claims Coverage
The independent claim identified in the provided material is directed to a method that alters the glycosylation pattern of a recombinant glycoprotein by using a cell culture medium supplemented with mycophenolic acid acyl glucuronide, with alteration defined relative to production using the same cells and the same medium without the additive. No other independent claims are included in the provided claim list.
Mycophenolic acid acyl glucuronide supplementation to alter glycosylation pattern
Culturing eukaryotic cells engineered to express a recombinant glycoprotein of interest in a cell culture medium, wherein the cell culture medium is supplemented with an additive comprising mycophenolic acid acyl glucuronide, and wherein the glycosylation pattern of the recombinant glycoprotein of interest is altered relative to the same recombinant glycoprotein produced by the same cells in the same cell culture medium without the additive.
Based on the provided independent claim, the claim coverage centers on the use of mycophenolic acid acyl glucuronide in a supplemented cell culture medium to alter the glycosylation pattern of a recombinant glycoprotein, explicitly defined by comparison to the same system without the additive.
Stated Advantages
Altering the glycosylation pattern of a recombinant glycoprotein relative to production in the same medium without the additive.
Modifying glycosylation attributes including afucosylation, galactosylation, α-gal, and/or N-glycolylneuraminic acid (NGNA).
Reducing FcγRIIIa binding and reducing antibody-dependent cell-mediated cytotoxicity (ADCC).
Changing glycosylation outcomes while addressing culture performance such as viability and titer.
Documented Applications
Producing recombinant glycoproteins in eukaryotic cell culture with controlled glycosylation patterns, including changes in afucosylation, galactosylation, α-gal, and NGNA, and associated functional glycan-related properties such as FcγRIIIa binding and ADCC.
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