Abstract
Nascent advanced therapies, including regenerative medicine and cell and
gene therapies, rely on the production of cells in bioreactors that are
highly heterogeneous in both space and time. Unfortunately, these
promising therapies have failed to reach a wide patient population due
to unreliable manufacturing processes that result in batch variability
and cost prohibitive production. This can be attributed largely to a
void in existing process analytical technologies (PATs) capable of
characterizing the secreted critical quality attributes (CQAs)
biomolecules that correlate with the final product quality. The Dynamic
Sampling Platform (DSP) is a PAT for cell bioreactor monitoring that can
be coupled to a suite of sensor techniques to provide real-time feedback
on spatial and temporal CQA content in situ. In this study, DSP is
coupled with electrospray ionization mass spectrometry (ESI-MS) and
direct-from-culture sampling to obtain measures of CQA content in bulk
media and the cell microenvironment throughout the entire cell culture
process (~3 weeks). Post hoc analysis of this real-time
data reveals that DSP output is heavily dependent on spatial context.
Importantly, these results demonstrate that an effective PAT must
incorporate both spatial and temporal resolution to serve as an
effective input f or feedback control in advanced therapy production.