Application of Raman spectroscopy during pharmaceutical process development for determination of critical quality attributes in Protein A chromatography

Chen, Jingyi; Wang, Jiarui; Hess, Rudger; Wang, Gang; Studts, Joey; Franzreb, Matthias

doi:10.1016/j.chroma.2024.464721

Cited by 2 publications

(1 citation statement)

References 23 publications

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“…Furthermore, by incorporating additional reference analytics such as HCP-enzyme-linked immunosorbent assay (ELISA) as done in Großhans et al (2019), the prediction of HCP content via Raman spectroscopic measurements may hence be facilitated. However, the prediction of HCPs using chemometric methods has only rarely been reported (Capito et al, 2015;Chen et al, 2024), most likely due to the diversity of species classified as HCPs (Falkenberg et al, 2019). The same applies to nucleic acids and endotoxins, which might affect VLP precipitation, but their analysis exceed the scope of this study.…”

Section: Effects Of Preprocessing Operations On Raman Datamentioning

confidence: 86%

Raman-based PAT for VLP precipitation: systematic data diversification and preprocessing pipeline identification

Dietrich,

Schiemer,

Kurmann

et al. 2024

Front. Bioeng. Biotechnol.

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Virus-like particles (VLPs) are a promising class of biopharmaceuticals for vaccines and targeted delivery. Starting from clarified lysate, VLPs are typically captured by selective precipitation. While VLP precipitation is induced by step-wise or continuous precipitant addition, current monitoring approaches do not support the direct product quantification, and analytical methods usually require various, time-consuming processing and sample preparation steps. Here, the application of Raman spectroscopy combined with chemometric methods may allow the simultaneous quantification of the precipitated VLPs and precipitant owing to its demonstrated advantages in analyzing crude, complex mixtures. In this study, we present a Raman spectroscopy-based Process Analytical Technology (PAT) tool developed on batch and fed-batch precipitation experiments of Hepatitis B core Antigen VLPs. We conducted small-scale precipitation experiments providing a diversified data set with varying precipitation dynamics and backgrounds induced by initial dilution or spiking of clarified Escherichia coli-derived lysates. For the Raman spectroscopy data, various preprocessing operations were systematically combined allowing the identification of a preprocessing pipeline, which proved to effectively eliminate initial lysate composition variations as well as most interferences attributed to precipitates and the precipitant present in solution. The calibrated partial least squares models seamlessly predicted the precipitant concentration with R2 of 0.98 and 0.97 in batch and fed-batch experiments, respectively, and captured the observed precipitation trends with R2 of 0.74 and 0.64. Although the resolution of fine differences between experiments was limited due to the observed non-linear relationship between spectral data and the VLP concentration, this study provides a foundation for employing Raman spectroscopy as a PAT sensor for monitoring VLP precipitation processes with the potential to extend its applicability to other phase-behavior dependent processes or molecules.

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