title: Near-infrared spectroscopic evaluation of lyophilized viral vaccine formulations.
abstract: This article examines the applicability of near-infrared spectroscopy (NIRS) to evaluate the virus state in a freeze-dried live, attenuated vaccine formulation. Therefore, this formulation was freeze-dried using different virus volumes and after applying different pre-freeze-drying virus treatments (resulting in different virus states): (i) as used in the commercial formulation; (ii) without antigen (placebo); (iii) concentrated via a centrifugal filter device; and (iv) stressed by 96 h exposure to room temperature. Each freeze-dried product was measured directly after freeze-drying with NIR spectroscopy and the spectra were analyzed using principal component analysis (PCA). Herewith, two NIR spectral regions were evaluated: (i) the 7300-4000 cm(-1) region containing the amide A/II band which might reflect information on the coated proteins of freeze-dried live, attenuated viruses; and (ii) the C-H vibration overtone regions (10,000-7500 and 6340-5500 cm(-1) ) which might supply information on the lipid layer surrounding the freeze-dried live, attenuated viruses. The different pre-freeze-drying treated live, attenuated virus formulations (different virus states and virus volumes) resulted in different clusters in the scores plots resulting from the PCA of the collected NIR spectra. Secondly, partial least squares discriminant analysis models (PLS-DA) were developed and evaluated, allowing classification of the freeze-dried formulations according to virus pretreatment. The results of this study suggest the applicability of NIR spectroscopy for evaluating live, attenuated vaccine formulations with respect to their virus pretreatment and virus volume.
authors.affiliation: Laboratory of Pharmaceutical Process Analytical Technology, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium.
authors.forename: Laurent
authors.initials: L
authors.lastname: Hansen
authors.forename: Thomas De
authors.initials: TD
authors.lastname: Beer
authors.forename: Sigrid
authors.initials: S
authors.lastname: Pieters
authors.forename: Yvan Vander
authors.initials: YV
authors.lastname: Heyden
authors.forename: Chris
authors.initials: C
authors.lastname: Vervaet
authors.forename: Jean Paul
authors.initials: JP
authors.lastname: Remon
authors.forename: Jean-Pierre
authors.initials: JP
authors.lastname: Montenez
authors.forename: Rim
authors.initials: R
authors.lastname: Daoussi
mesh_terms: D002626:Chemistry, Pharmaceutical; D005612:Freeze Drying; D006801:Humans; D019265:Spectroscopy, Near-Infrared; D014613:Vaccines, Attenuated / Q000737:chemistry* / Q000627:therapeutic use; D014765:Viral Vaccines / Q000737:chemistry* / Q000627:therapeutic use
chemical_list: D014613:Vaccines, Attenuated; D014765:Viral Vaccines
keywords: NIR spectroscopy; attenuated vaccines; freeze drying; live; partial least square discriminant analysis (PLSDA); principal component analysis (PCA); process analytical technology (PAT)