Mammalian cells exhibit inefficient and poorly regulated metabolism as they tend to use as much as the available nutrients and produce waste metabolites rather than the required amount for their growth. As a result, they produce substantial waste products that create a hostile environment for the growth of cells as well as the target protein and lowers the protein quality. In order to control the metabolism and increase the productivity, it’s crucial to identify the inhibitory waste metabolites and study their pathway. The work flow for the waste inhibitor metabolomics are candidate identification, inhibitory impact verification and quantification, and control strategy development. In this study, 6 novel inhibitory metabolites were identified based on their impact on growth, productivity, and product quality. The metabolome profiles of VRC O1 (CHO K1) cell line was firstly screening in High Cell Density fed-batch cultures using high resolution global metabolomics platforms, Orbitrap LC-MS. The top 18 inhibitor candidates with high confidence in identification matching were selected by multivariate data analysis and pathway analysis. The high ranked candidates were experimentally verified by spiking them in the batch culture. Among them, 6 metabolites inhibit growth, 6 metabolites lower the antibody production, and 8 metabolites show impact on IgG glycosylation.

Identifying inhibitory waste by-products in high density CHO cell culture

Macromolecular Chemistry: The Second Century

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ACS Spring 2020 National Meeting & Expo

The official archive and repository for all ACS Meetings & Expositions abstracts and select posters or presentations.