SYSTEMS BIOTECHNOLOGY TOOLS FOR GUIDING THE DEVELOPMENT OF CHO CELL BIOPROCESSING

Abstract

While empirical approaches have been used to improve Chinese hamster ovary (CHO) cell bioprocessing for biopharmaceutical production, they are largely done without a holistic and mechanistic perspective of its metabolism that is necessary for informing strategies to improve performance. To do so, in silico tools have been developed in this thesis. The first is a pre-processing pipeline for the efficient and accurate profiling of the CHO lipidome, based on LC-MS platform. The second tool is a lipid table for efficiently discerning species levels, metabolism trends, and their effects on the bioprocess. Enzyme constraints -based flux balance analysis (ec-FBA) is further developed as a third tool for improving flux predictions in CHO cells. The last tool combines partial-least-square regression analysis with ec-FBA for the systematic and rational reformulation of cell culture medium for enhancing growth and productivity. Thus, the tools allow for a more efficient and effective development of CHO cell bioprocessing.