Synchronous fluorescence spectroscopy as a novel tool to enable PAT applications in bioprocesses

Area of Science:

• Biotechnology
• Analytical Chemistry
• Process Analytical Technology (PAT)

Background:

• Traditional 2D fluorometry faces limitations in speed and resolution for bioprocess monitoring.
• Real-time monitoring is crucial for optimizing animal cell culture processes and ensuring product quality.

Purpose of the Study:

• To evaluate synchronous fluorescence spectroscopy (SFS) as a novel tool for in situ, real-time monitoring of animal cell cultures.
• To compare the performance of SFS against traditional 2D fluorometry for monitoring cell density and antibody titer.

Main Methods:

• In situ monitoring of CHO cell cultures using both 2D and SFS techniques.
• Development of partial least squares regression models for viable cell density and antibody levels.
• Validation of predictive models using a held-out portion of the dataset.

Main Results:

• SFS provided improved peak resolution and recording speed compared to 2D fluorometry.
• Both techniques contained sufficient information for distinguishing cell cultures via principal component analysis.
• SFS models achieved high validation accuracies (>91%) for predicting cell density and antibody titer.

Conclusions:

• SFS is a superior and faster alternative to 2D fluorometry for real-time bioprocess monitoring.
• SFS facilitates enhanced process control strategies within a Process Analytical Technology (PAT) framework.
• This technique enables efficient tracking of cell growth and product formation in bioreactors.