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Modulation of Antibody N-Linked Glycosylation in Dynamic Perfusion CHO Cell Cultures by Mn 2+ Supplementation
  • Sam Preza,
  • Matthew Radle,
  • Ken Lee
Sam Preza
AstraZeneca R&D Gaithersburg

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Matthew Radle
AstraZeneca R&D Gaithersburg
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Ken Lee
AstraZeneca R&D Gaithersburg
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Abstract

Companies across the biotechnology industry have been investing in the development of cell culture perfusion technologies. Perfusion, part of the Next Generation of Manufacturing (NGM) platforms, has demonstrated dramatic increases in volumetric productivity of therapeutic proteins compared to traditional fed-batch processes. There are typically two modes of operation for perfusion cell cultures: steady state and dynamic. Steady state perfusion commonly employs a cell bleed to maintain manageable viable cell density (VCD) targets. In contrast, dynamic perfusion allows for undisturbed cell growth which leads to higher cell densities and total productivity. The trade-off comes at an increased engineering complexity required to ensure cell health and maintain target critical quality attributes (CQAs) at higher cell densities. Specifically, N-linked glycosylation is a CQA for the NGM upstream platform due to its potential role in a protein’s pharmacokinetics (PK) and pharmacodynamics (PD). In this study we show how control of dynamic Mn 2+ bioreactor supplementation allows for consistent production of desired glycosylation profiles in a dynamic perfusion mode. We utilized a design of experiment (DoE) to test the effect of dynamic Mn 2+ supplementation on dynamic and steady state perfusion. It was found that Mn 2+ supplementation inhibits cell growth above 2 µM but successfully maintained desired glycosylation targets.