Use of an Inducible CHO System to Research Metabolism and Recombinant Therapeutic Glycoprotein Production

Abstract

Glycosylated recombinant therapeutic proteins (GRTPs) are important drugs for the treatment of cancers and autoimmune disorders, but a single formulation contains structures that are highly heterogeneous and dependent on cellular metabolism. Protein structure translates to product quality; therefore, a key goal of industry is to minimise heterogeneity, and produce only structures with preferable characteristics. Sugars attached to proteins (glycans) are a major source of this heterogeneity. Chinese hamster ovary (CHO) cells are a leading platform for producing GRTPs. Cellular metabolism links the culture environment to the product, but its role is not yet fully understood, as the interaction is comprised of a highly complex network of events. Recent research efforts have provided insight into these networks, but there is scope to investigate this further, the understanding of which can improve process efficiency. Our work shows development of a glycan analytical method at Manchester University for GRTP structural elucidation. An inducible CHO cell line was also created, tailored specifically for understanding metabolic networks involved in GRTP production. Finally, this cell line was tested with a cellular feeding strategy, providing useful insight into cell culture environment considerations for cell lines producing GRTPs. This research provides a toolbox to simplify research into the role of metabolism in CHO cell environment and GRTP production. This has implications for future cell culture feed design and metabolic engineering strategies.

Date of Award	31 Dec 2017
Original language	English
Awarding Institution	The University of Manchester
Supervisor	Ewan Blanch (Supervisor) & Alan Dickson (Supervisor)