Open AccessModel-Driven Engineering of N-Linked Glycosylation in Chinese Hamster Ovary Cells
Author(s) -
Christopher S. Stach,
Meghan G. McCann,
Conor M. O'Brien,
Tung Le,
Nikunj V. Somia,
Xinning Chen,
Kyung Ho Lee,
Hsu Yuan Fu,
Pródromos Daoutidis,
Liang Zhao,
Wei Shou Hu,
Michael J. Smanski
Publication year - 2019
Publication title -
acs synthetic biology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.156
H-Index - 66
ISSN - 2161-5063
DOI - 10.1021/acssynbio.9b00215
Subject(s) - chinese hamster ovary cell , glycosylation , glycan , computational biology , biology , synthetic biology , transgene , systems biology , gene , microbiology and biotechnology , glycoprotein , cell culture , biochemistry , genetics
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.