Fluorescence spectroscopy in combination with multivariate statistical methods was employed as a tool for monitoring the manufacturing process of pertactin (PRN), one of the virulence factors of Bordetella pertussis utilized in whopping cough vaccines. Fluorophores such as amino acids and co-enzymes were detected throughout the process. The fluorescence data collected at different stages of the fermentation and purification process were treated employing principal component analysis (PCA). Through PCA, it was feasible to identify sources of variability in PRN production. Then, partial least square (PLS) was employed to correlate the fluorescence spectra obtained from pure PRN samples and the final protein content measured by a Kjeldahl test from these samples. In view that a statistically significant correlation was found between fluorescence and PRN levels, this approach could be further used as a method to predict the final protein content.
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The authors would like to thank NSERC (National Science and Engineering Research Council of Canada) and Sanofi Pasteur for the financial support.
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Zavatti, V., Budman, H., Legge, R. et al. Monitoring of an antigen manufacturing process. Bioprocess Biosyst Eng 39, 855–869 (2016). https://doi.org/10.1007/s00449-016-1565-1
- Principal component analysis
- Partial least square
- Fluorescence spectroscopy
- Antigen manufacturing
- Bioprocess monitoring
- Bordetella pertussis