Laboratory of Pharmaceutical Process Analytical Technology, Ghent University, Harelbekestraat 72, 9000, Ghent, Belgium.
Biotechnol Prog. 2013 Nov-Dec;29(6):1573-86. doi: 10.1002/btpr.1807. Epub 2013 Sep 30.
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.
本文研究了近红外光谱（NIRS）在评估冻干减毒活疫苗制剂中病毒状态的适用性。因此，该制剂采用不同的病毒量进行冷冻干燥，并在应用不同的预冷冻干燥病毒处理（导致不同的病毒状态）后进行冷冻干燥：（i）用于商业制剂；（ii）无抗原（安慰剂）；（iii）通过离心过滤装置浓缩；（iv）在室温下暴露 96 小时以进行应激处理。每个冻干产品在冷冻干燥后直接用 NIR 光谱进行测量，并使用主成分分析（PCA）对光谱进行分析。在此，评估了两个 NIR 光谱区域：（i）包含酰胺 A/II 带的 7300-4000 cm(-1) 区域，该区域可能反映冻干减毒活病毒包被蛋白的信息；（ii）C-H 振动倍频区（10000-7500 和 6340-5500 cm(-1) ），可能提供冻干减毒活病毒周围脂质层的信息。不同的预冷冻干燥处理的减毒活病毒制剂（不同的病毒状态和病毒量）导致 PCA 收集的 NIR 光谱的得分图中出现不同的聚类。其次，开发并评估了偏最小二乘判别分析模型（PLS-DA），允许根据病毒预处理对冻干制剂进行分类。该研究的结果表明，NIR 光谱可用于评估减毒活疫苗制剂的病毒预处理和病毒量。
