Modulation of Antibody N-Linked Glycosylation in Dynamic Perfusion CHO
Cell Cultures by Mn 2+ Supplementation
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.