Perfusion cell culture offers attractive benefits over fed batch but comes with its own set of challenges. To maintain steady state cell density, a volume of cell culture needs to be removed daily, creating in essence a ‘cell bleed’. For fast growing cell lines, the product loss from the cell bleed stream can be upwards of 30%, significantly impacting process efficiency. The concept of two phases of perfusion cell culture is therefore presented here: a fast growing ramp up phase followed by a slow growing production phase. The ability to control cell growth thus becomes crucial. Leveraging existing media components to achieve such control without introducing additional supplements is most desirable because it decreases process complexity and eliminates safety and clearance concerns.<br/><br/>Media components were identified that impact the cell growth signaling cascade. Altering concentrations of specific media components effectively impacted intracellular chemistry as determined by flow cytometry. A state of cell cycle arrest was subsequently induced. Cell cycle analysis revealed a significantly higher percentage of cells in the G0/1 state, which had previously been reported by other groups as being the most productive state. In alignment with those previous reports, cell specific productivities were drastically increased by as much as 4-fold in these arrested cells, increasing the productivity from 30pg/cell/day to as much as 115pg/cell/day in one case. The results for multiple cell lines and the impact on product quality attributes will be discussed.

Leveraging media components to control cell growth and improve specific productivity in perfusion cell culture

253rd National Meeting (2017)

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