In order to shorten and simplify the current upstream seed train culture expansion operations, a perfusion-based, non-centrifugal high density (HD) cell banking platform was developed. Various cell retention methods including alternating tangential flow (ATF, by Refine Technologies), tangential flow filtration (TFF), and built-in floating filter were evaluated. ATF4 was selected because of its superior performance in supporting a HD perfusion culture in a 10-L Wave Cellbag bioreactor as well as a fast in-vessel cell concentration.  A simplified process control strategy was also developed so that desired pH and DO ranges can be maintained by manually adjusting gas compositions and rocking conditions. This strategy eliminated the dependence on online pH and DO feedback controls. When the culture reached a viable cell density of > 3�107/mL, it was concentrated to > 11�107/mL, and was then frozen at 10�107/mL in 5-mL cryovials. It has been demonstrated that one HD vial enables the direct inoculation of a 1 L culture with high growth rate and viability. This HD banking platform has been successfully tested with three CHO cell lines expressing different recombinant proteins/antibodies.

Development of a perfusion-based non-centrifugal high density cell banking platform

247th National Meeting (2014)

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