title: Cell culture media supplementation of uncommonly used sugars sucrose and tagatose for the targeted shifting of protein glycosylation profiles of recombinant protein therapeutics.
abstract: Protein glycosylation is an important post-translational modification toward the structure and function of recombinant therapeutics. The addition of oligosaccharides to recombinant proteins has been shown to greatly influence the overall physiochemical attributes of many proteins. It is for this reason that protein glycosylation is monitored by the developer of a recombinant protein therapeutic, and why protein glycosylation is typically considered a critical quality attribute. In this work, we highlight a systematic study toward the supplementation of sucrose and tagatose into cell culture media for the targeted modulation of protein glycosylation profiles on recombinant proteins. Both sugars were found to affect oligosaccharide maturation resulting in an increase in the percentage of high mannose N-glycan species, as well as a concomitant reduction in fucosylation. The latter effect was demonstrated to increase antibody-dependent cell-mediated cytotoxicity for a recombinant antibody. These aforementioned results were found to be reproducible at different scales, and across different Chinese hamster ovary cell lines. Through the selective supplementation of these described sugars, the targeted modulation of protein glycosylation profiles is demonstrated, as well as yet another tool in the cell culture toolbox for ensuring product comparability.
authors.affiliation: Process Sciences-Cell Culture, AbbVie Inc., Worcester, MA, 01605.
authors.forename: Patrick
authors.initials: P
authors.lastname: Hossler
authors.forename: Sean
authors.initials: S
authors.lastname: McDermott
authors.forename: Christopher
authors.initials: C
authors.lastname: Racicot
authors.forename: Christopher
authors.initials: C
authors.lastname: Chumsae
authors.forename: Haly
authors.initials: H
authors.lastname: Raharimampionona
authors.forename: Yu
authors.initials: Y
authors.lastname: Zhou
authors.forename: David
authors.initials: D
authors.lastname: Ouellette
authors.forename: Joseph
authors.initials: J
authors.lastname: Matuck
authors.forename: Ivan
authors.initials: I
authors.lastname: Correia
authors.forename: John
authors.initials: J
authors.lastname: Fann
authors.forename: Jianmin
authors.initials: J
authors.lastname: Li
mesh_terms: D000818:Animals; D000906:Antibodies / Q000032:analysis / Q000737:chemistry / Q000378:metabolism; D016466:CHO Cells; D018929:Cell Culture Techniques / Q000379:methods*; D006224:Cricetinae; D003412:Cricetulus; D003470:Culture Media / Q000737:chemistry* / Q000378:metabolism; D006031:Glycosylation; D006601:Hexoses / Q000737:chemistry / Q000378:metabolism*; D011994:Recombinant Proteins / Q000032:analysis / Q000737:chemistry* / Q000378:metabolism*; D013395:Sucrose / Q000737:chemistry / Q000378:metabolism*
chemical_list: D000906:Antibodies; D003470:Culture Media; D006601:Hexoses; D011994:Recombinant Proteins; D013395:Sucrose; C030192:tagatose
keywords: mammalian cell culture; product quality; protein glycosylation; sucrose; tagatose