2017 Annual Meeting
(67a) Modifying IgG Glycan Profile Using a High-Throughput Multi-Gene Assembly Platform
Authors
Christopher Stach - Presenter, University of Minnesota, Twin Cities
Wei-Shou Hu, University of Minnesota, Twin Cities
Michael J. Smanski, University of Minnesota, Twin Cities
Tung S. Le, University of Minnesota, Twin Cities
Meghan G. McCann, University of Minnesota, Twin Cities
Liang Zhao, East China University of Science & Technology
Xinning Chen, East China University of Science & Technology
Protein N-glycosylation is a post-translational modification that is crucial for protein folding, biological activity, half-life and stability. It is one of the most complex post-translational modifications in that it involves a complex network of enzymes that may act on identical substrates to generate many different glycosylation patterns. A drawback of protein expression in mammalian cells is the presence of heterogeneous populations of the same protein due to differences in protein N-glycosylation. The complex nature of glycosylation makes it a good target for control using genetic engineering approaches. While pipelines exist for the generation of multi-gene constructs they are limited in their utility. Utilizing the latest DNA synthesis and assembly technology we developed a platform for combinatorial assembly of multi-gene constructs for use in altering glycosylation of immunoglobulin G (IgG) produced in chinese hamster ovary cells (CHO). A kinetic based model of N-glycosylation was used to predict N-glycosylation genes responsible for generating targeted glycosylation profiles of IgG. Expression of selected glycosylation genes from multi-gene constructs led to altered glycan profiles of IgG demonstrating the efficacy of our platform and validating predictions of our model.