Monitoring small-scale bioreactor studies for media development using polarized total synchronous fluorescence...

Area of Science:

• Biopharmaceutical Process Development
• Spectroscopic Analysis
• Bioprocess Monitoring

Background:

• Small-scale bioreactors (SSBR) are crucial for optimizing media and conditions in biopharmaceutical production.
• Key process parameters (CPP) like protein titre and viable cell density (VCD) require accurate monitoring during SSBR studies.

Purpose of the Study:

• To evaluate polarized total synchronous fluorescence spectroscopy (TSFS||) and Synchronous Light Scattering (SyLS||) for monitoring CPPs in SSBR.
• To quantitatively predict protein titre and VCD using spectroscopic methods during media optimization.

Main Methods:

• Utilized TSFS|| and SyLS|| on samples from a 13-day SSBR study with 71 media formulations.
• Applied statistical analysis, principal component analysis (PCA), and Support Vector Machines (SVM) for data interpretation.
• Correlated spectroscopic data with particle size from Dynamic Light Scattering.

Main Results:

• TSFS|| spectra showed significant changes correlating with bioprocess progression.
• SyLS|| correlated with particle size but not VCD due to sample preparation.
• Spectral variations were higher between media formulations than process evolution, limiting global prediction models.
• Classification methods successfully identified media subsets for improved prediction accuracy.

Conclusions:

• TSFS|| and SyLS|| offer a holistic, multi-attribute approach to bioprocess monitoring, reducing reliance on offline methods.
• Classification models, like SVM, are effective for screening media formulations in SSBR.
• This spectroscopic approach can inform in-process monitoring strategies for production-scale bioprocesses.