Model-Driven Engineering of N-Linked Glycosylation in Chinese Hamster Ovary Cells

Christopher S. Stach, Meghan G. McCann, Conor M. O'Brien, Tung S. Le, Nikunj Somia, Xinning Chen, Kyoungho Lee, Hsu Yuan Fu, Prodromos Daoutidis, Liang Zhao, Wei Shou Hu, Michael Smanski

Research output: Contribution to journalArticlepeer-review

13 Scopus citations

Abstract

Chinese hamster ovary (CHO) cells are used for industrial production of protein-based therapeutics (i.e., "biologics"). Here we describe a method for combining systems-level kinetic models with a synthetic biology platform for multigene overexpression to rationally perturb N-linked glycosylation. Specifically, we sought to increase galactose incorporation on a secreted Immunoglobulin G (IgG) protein. We rationally design, build, and test a total of 23 transgenic cell pools that express single or three-gene glycoengineering cassettes comprising a total of 100 kilobases of engineered DNA sequence. Through iterative engineering and model refinement, we rationally increase the fraction of bigalactosylated glycans five-fold from 11.9% to 61.9% and simultaneously decrease the glycan heterogeneity on the secreted IgG. Our approach allows for rapid hypothesis testing and identification of synergistic behavior from genetic perturbations by bridging systems and synthetic biology.

Original languageEnglish (US)
Pages (from-to)2524-2535
Number of pages12
JournalACS Synthetic Biology
Volume8
Issue number11
DOIs
StatePublished - Nov 15 2019

Bibliographical note

Publisher Copyright:
© 2019 American Chemical Society.

Keywords

  • CHO cells
  • DNA assembly
  • IgG glycosylation
  • post-translational modification
  • systems-level modeling

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