A Novel Approach Using Real-time Dynamic Imaging Analysis to Monitor
Cellular Apoptosis, Viability, and Cell Density in CHO Cell-based
Bioprocesses for Monoclonal Antibody Production.
Abstract
The biopharmaceutical industry continually seeks innovative approaches
to optimize monoclonal antibody (mAb) production, aligning with patient
demands, cost-effectiveness, and revenue growth. Chinese hamster ovary
(CHO) cells play a pivotal role in mAb production. However, undesired
massive CHO cell death can adversely impact productivity, necessitating
preventive control operations. To ensure effective control and mitigate
the negative impact of cell death, real-time or near real-time
monitoring of cell health status is crucial. Bioprocess operators can
promptly identify deviations from the desired cell status and take
appropriate actions to maintain optimal conditions for mAb production.
Additionally, in-process cell status information enhances process
understanding, providing valuable insights into the underlying
mechanisms governing cell behavior and maximizing productivity. In this
study, we investigated an innovative approach, Canty
TM dynamic imaging analysis (DIA), as a promising
solution for real-time monitoring of cellular apoptosis, viability, and
cell density. The conventional trypan blue exclusion method (Vi-CELL
TM) and To-Pro-3 fluorescent dye staining (flow
cytometry) were used as standard reference methods. Our findings
revealed that the Canty TM DIA method combined with
appropriate mathematical modeling yielded results comparable to those
obtained from Vi-CELL TM and flow cytometry methods.
Canty TM DIA successfully traced the trajectory of
cell death progress, detecting the onset of apoptosis earlier than the
following cell death event. This early detection capability would allow
timely intervention to prevent further cell death and maintain optimal
productivity. Furthermore, Canty TM DIA demonstrated
superior performance in distinguishing images of aggregated cells,
providing a more accurate measure of total cell density. This improved
accuracy is crucial for calculating viable cell density, which reflects
the overall health status of the cells. The results highlight the
potential of Canty TM DIA as a powerful tool for
real-time monitoring of cell status throughout mAb production processes.