title: A dynamic mathematical model for monoclonal antibody N-linked glycosylation and nucleotide sugar donor transport within a maturing Golgi apparatus.
abstract: Monoclonal antibodies (mAbs) are one of the most important products of the biopharmaceutical industry. Their therapeutic efficacy depends on the post-translational process of glycosylation, which is influenced by manufacturing process conditions. Herein, we present a dynamic mathematical model for mAb glycosylation that considers cisternal maturation by approximating the Golgi apparatus to a plug flow reactor and by including recycling of Golgi-resident proteins (glycosylation enzymes and transport proteins [TPs]). The glycosylation reaction rate expressions were derived based on the reported kinetic mechanisms for each enzyme, and transport of nucleotide sugar donors [NSDs] from the cytosol to the Golgi lumen was modeled to serve as a link between glycosylation and cellular metabolism. Optimization-based methodologies were developed for estimating unknown enzyme and TP concentration profile parameters. The resulting model is capable of reproducing glycosylation profiles of commercial mAbs. It can further reproduce the effect gene silencing of the FucT glycosylation enzyme and cytosolic NSD depletion have on the mAb oligosaccharide profile. All novel elements of our model are based on biological evidence and generate more accurate results than previous reports. We therefore believe that the improvements contribute to a more detailed representation of the N-linked glycosylation process. The overall results show the potential of our model toward evaluating cell engineering strategies that yield desired glycosylation profiles. Additionally, when coupled to cellular metabolism, this model could be used to assess the effect of process conditions on glycosylation and aid in the design, control, and optimization of biopharmaceutical manufacturing processes.
authors.affiliation: Dept. of Chemical Engineering and Chemical Technology, Imperial College London, South Kensington Campus, London, UK.
authors.forename: Ioscani
authors.initials: I
authors.lastname: Jimenez del Val
authors.forename: Judit M
authors.initials: JM
authors.lastname: Nagy
authors.forename: Cleo
authors.initials: C
authors.lastname: Kontoravdi
mesh_terms: D020816:Amino Acid Motifs; D000818:Animals; D000911:Antibodies, Monoclonal / Q000737:chemistry / Q000378:metabolism*; D001692:Biological Transport; D002460:Cell Line; D004798:Enzymes / Q000737:chemistry / Q000378:metabolism; D006031:Glycosylation; D006056:Golgi Apparatus / Q000737:chemistry / Q000201:enzymology / Q000378:metabolism*; D006801:Humans; D007700:Kinetics; D008962:Models, Theoretical; D009702:Nucleoside Diphosphate Sugars / Q000378:metabolism*; D011499:Protein Processing, Post-Translational
chemical_list: D000911:Antibodies, Monoclonal; D004798:Enzymes; D009702:Nucleoside Diphosphate Sugars