title: Arabinosylation of recombinant human immunoglobulin-based protein therapeutics.
abstract: Protein glycosylation is arguably the paramount post-translational modification on recombinant glycoproteins, and highly cited in the literature for affecting the physiochemical properties and the efficacy of recombinant glycoprotein therapeutics. Glycosylation of human immunoglobulins follows a reasonably well-understood metabolic pathway, which gives rise to a diverse range of asparagine-linked (N-linked), or serine/threonine-linked (O-linked) glycans. In N-linked glycans, fucose levels have been shown to have an inverse relationship with the degree of antibody-dependent cell-mediated cytotoxicity, and high mannose levels have been implicated in potentially increasing immunogenicity and contributing to less favorable pharmacokinetic profiles. Here, we demonstrate a novel approach to potentially reduce the presence of high-mannose species in recombinant human immunoglobulin preparations, as well as facilitate an approximate 100% replacement of fucosylation with arabinosylation in Chinese hamster ovary cell culture through media supplementation with D-arabinose, an uncommonly used mammalian cell culture sugar substrate. The replacement of fucose with arabinose was very effective and practical to implement, since no cell line engineering or cellular adaptation strategies were required. Arabinosylated recombinant IgGs and the accompanying reduction in high mannose glycans, facilitated a reduction in dendritic cell uptake, increased FcγRIIIa signaling, and significantly increased the levels of ADCC. These aforementioned effects were without any adverse changes to various structural or functional attributes of multiple recombinant human antibodies and a bispecific DVD-Ig. Protein arabinosylation represents an expansion of the N-glycan code in mammalian expressed glycoproteins.
authors.affiliation: a Process Sciences, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Patrick
authors.initials: P
authors.lastname: Hossler
authors.affiliation: a Process Sciences, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Chumsae
authors.affiliation: a Process Sciences, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Christopher
authors.initials: C
authors.lastname: Racicot
authors.affiliation: a Process Sciences, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: David
authors.initials: D
authors.lastname: Ouellette
authors.affiliation: b Preclinical Safety, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Alexander
authors.initials: A
authors.lastname: Ibraghimov
authors.affiliation: b Preclinical Safety, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Daniel
authors.initials: D
authors.lastname: Serna
authors.affiliation: a Process Sciences, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Mora
authors.affiliation: a Process Sciences, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Sean
authors.initials: S
authors.lastname: McDermott
authors.affiliation: c Discovery-Biologics, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Boris
authors.initials: B
authors.lastname: Labkovsky
authors.affiliation: b Preclinical Safety, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Susanne
authors.initials: S
authors.lastname: Scesney
authors.affiliation: d Drug Metabolism and Pharmacokinetics, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Christine
authors.initials: C
authors.lastname: Grinnell
authors.affiliation: d Drug Metabolism and Pharmacokinetics, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Gregory
authors.initials: G
authors.lastname: Preston
authors.affiliation: c Discovery-Biologics, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Sahana
authors.initials: S
authors.lastname: Bose
authors.affiliation: e Preformulation, AbbVie Bioresearch Center , Worcester , MA , USA.
authors.forename: Ralf
authors.initials: R
authors.lastname: Carrillo
mesh_terms: D000818:Animals; D000911:Antibodies, Monoclonal / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D000920:Antibody-Dependent Cell Cytotoxicity; D001089:Arabinose / Q000494:pharmacology*; D016466:CHO Cells; D003412:Cricetulus; D006031:Glycosylation / Q000187:drug effects; D006801:Humans; D007074:Immunoglobulin G / Q000097:blood* / Q000737:chemistry / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis / Q000737:chemistry / Q000235:genetics
chemical_list: D000911:Antibodies, Monoclonal; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D001089:Arabinose
keywords: ADCC; Fc effector activity; afucosylation; arabinose; protein glycosylation