Fed-batch bioreactors are the current industry standard for production using CHO cells due to ease of scale-up and compatibility with large-scale manufacturing. In fed-batch microbial cultures, nutrients are continuously or semi-continuously fed, while the effluent may be removed and if so, discontinuously. Typically, fed-batch culture conditions are used to adjust metabolic rates and overcome substrate inhibition by controlled substrate feeding. Currently, most CHO cell culture fed-batch feeding strategies utilize daily bolus feeding. This discontinuous feeding approach often relies on off-line daily measurement of a key substrate (e.g., glucose) that targets to maintain this metabolite at or above a desired concentration. With daily bolus feeding, cells are typically not limited in growth and, therefore, exhibit an unrestricted exponential growth rate. By establishing an exponential feeding profile, a desired growth rate could be achieved by maintaining CHO cells in a state of substrate limitation according to a predetermined desired growth rate, much like already commonly practiced for microbial fermentations. Preliminary data shows IgG production increases as cell growth begins to slow. A lower controlled growth rate may allow for redirection of cellular resources to protein production and reduce the burden of oxygen transport at high cell densities. Thus, it was hypothesized that slowing the growth rate earlier in the fed-batch phase for CHO cells would result in increased IgG production by allowing cells to redirect cellular resources to IgG production for a longer duration and ultimately result in higher IgG concentrations with improved protein quality. Pumping systems within the ambr®250 were used to control glucose feeding to achieve desired submaximal exponential cell growth rates. The results of this study show the impact of controlled glucose feeding on the growth profiles of CHO cells in fed-batch cultures and protein productivity.

Identifying glucose feeding profiles to control growth rate in CHO cells using precision pumping in the ambr®250 bioreactor system

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.