title: Monitoring of an antigen manufacturing process.
abstract: 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.
authors.affiliation: Department of Chemical Engineering, University of Waterloo, Waterloo, ON, Canada.
authors.forename: Vanessa
authors.initials: V
authors.lastname: Zavatti
authors.affiliation: Department of Chemical Engineering, University of Waterloo, Waterloo, ON, Canada. hbudman@uwaterloo.ca.
authors.forename: Hector
authors.initials: H
authors.lastname: Budman
authors.affiliation: Department of Chemical Engineering, University of Waterloo, Waterloo, ON, Canada.
authors.forename: Raymond
authors.initials: R
authors.lastname: Legge
authors.affiliation: Sanofi Pasteur, Toronto, ON, Canada.
authors.forename: Melih
authors.initials: M
authors.lastname: Tamer
mesh_terms: D000942:Antigens, Bacterial / Q000737:chemistry*; D001886:Bordetella pertussis / Q000276:immunology; D025341:Principal Component Analysis; D013050:Spectrometry, Fluorescence
chemical_list: D000942:Antigens, Bacterial
keywords: Antigen manufacturing; Bioprocess monitoring; Bordetella pertussis; Fluorescence spectroscopy; Partial least square; Principal component analysis; Vaccines