文献类型：期刊文献

英文题名：Model-Driven Engineering of N-Linked Glycosylation in Chinese Hamster Ovary Cells

作者：Stach, Christopher S.[1,2];McCann, Meghan G.[3];O'Brien, Conor M.[3];Le, Tung S.[3];Somia, Nikunj[4];Chen, Xinning[5];Lee, Kyoungho[3];Fu, Hsu-Yuan[3];Daoutidis, Prodromos[3];Zhao, Liang[5];Hu, Wei-Shou[3];Smanski, Michael[1,2]

机构：[1]Univ Minnesota, Dept Biochem Mol Biol & Biophys, Minneapolis, MN 55455 USA;[2]Univ Minnesota, Biotechnol Inst, Minneapolis, MN 55455 USA;[3]Univ Minnesota, Dept Chem Engn & Mat Sci, 421 Washington Ave SE, Minneapolis, MN 55455 USA;[4]Univ Minnesota, Dept Genet Cell Biol & Dev, Minneapolis, MN 55455 USA;[5]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2019

卷号：8

期号：11

起止页码：2524

外文期刊名：ACS SYNTHETIC BIOLOGY

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000497263400009)】；

基金：C.S.S. and M.G.M. are supported in part by a grant from the University of Minnesota Office of the Vice President of Research. M.G.M. is supported in part by the NIGMS Biotechnology Training Program (T32GM008347-22). MJ.S. is supported by The Defense Advanced Research Projects Agency (Grant No. D17AP00028).

语种：英文

外文关键词：IgG glycosylation; CHO cells; DNA assembly; systems-level modeling; post-translational modification

摘要：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.