title: N-linked glycosylation is an important parameter for optimal selection of cell lines producing biopharmaceutical human IgG.
abstract: We studied the variations in N-linked glycosylation of human IgG molecules derived from 105 different stable cell lines each expressing one of the six different antibodies. Antibody expression was based on glutamine synthetase selection technology in suspension growing CHO-K1SV cells. The glycans detected on the Fc fragment were mainly of the core-fucosylated complex type containing zero or one galactose and little to no sialic acid. The glycosylation was highly consistent for the same cell line when grown multiple times, indicating the robustness of the production and glycan analysis procedure. However, a twofold to threefold difference was observed in the level of galactosylation and/or non-core-fucosylation between the 105 different cell lines, suggesting clone-to-clone variation. These differences may change the Fc-mediated effector functions by such antibodies. Large variation was also observed in the oligomannose-5 glycan content, which, when present, may lead to undesired rapid clearance of the antibody in vivo. Statistically significant differences were noticed between the various glycan parameters for the six different antibodies, indicating that the variable domains and/or light chain isotype influence Fc glycosylation. The glycosylation altered when batch production in shaker was changed to fed-batch production in bioreactor, but was consistent again when the process was scaled from 400 to 5,000 L. Taken together, the observed clone-to-clone glycosylation variation but batch-to-batch consistency provides a rationale for selection of optimal production cell lines for large-scale manufacturing of biopharmaceutical human IgG.
authors.affiliation: Genmab and Immunotherapy Laboratory, Dept. of Immunology, University Medical Center, Utrecht, The Netherlands. p.vanberkel@genmab.com
authors.forename: Patrick H C
authors.initials: PH
authors.lastname: van Berkel
authors.forename: Jolanda
authors.initials: J
authors.lastname: Gerritsen
authors.forename: Gerrard
authors.initials: G
authors.lastname: Perdok
authors.forename: Jesper
authors.initials: J
authors.lastname: Valbjørn
authors.forename: Tom
authors.initials: T
authors.lastname: Vink
authors.forename: Jan G J
authors.initials: JG
authors.lastname: van de Winkel
authors.forename: Paul W H I
authors.initials: PW
authors.lastname: Parren
mesh_terms: D000818:Animals; D016466:CHO Cells; D002460:Cell Line / Q000166:cytology / Q000378:metabolism*; D006224:Cricetinae; D003412:Cricetulus; D006031:Glycosylation; D006801:Humans; D007074:Immunoglobulin G / Q000096:biosynthesis*; D011134:Polysaccharides / Q000378:metabolism
chemical_list: D007074:Immunoglobulin G; D011134:Polysaccharides