High-Throughput Raman Spectroscopy Combined with Innovate Data Analysis Workflow to Enhance Biopharmaceutical Process Development

Processes

Matthew Cheeks¹  Chris Spencer¹  Karolina Les²  Alexandra Umprecht²  Alison Tang²  Richard Turner²  Aled Charles²  Martyn Hulley²  Roman Zakrzewski³  SuzanneS. Farid³  Stephen Goldrick³ 

[1] Cell Culture & Fermentation Science, Biopharmaceuticals Development, R&D, AstraZeneca, Cambridge CB21 6GH, UK;Purification Process Sciences, Biopharmaceuticals Development, R&D, AstraZeneca, Cambridge CB21 6GH, UK;The Advanced Centre of Biochemical Engineering, Department of Biochemical Engineering, University College London, Gordon Street, London WC1E 6BT, UK;

关键词: Raman spectroscopy;    mammalian cell culture;    process analytical technology;    high-throughput;    scale-down technologies;    cation exchange chromatography;   

DOI  :  10.3390/pr8091179

来源: DOAJ

【 摘 要 】

Raman spectroscopy has the potential to revolutionise many aspects of biopharmaceutical process development. The widespread adoption of this promising technology has been hindered by the high cost associated with individual probes and the challenge of measuring low sample volumes. To address these issues, this paper investigates the potential of an emerging new high-throughput (HT) Raman spectroscopy microscope combined with a novel data analysis workflow to replace off-line analytics for upstream and downstream operations. On the upstream front, the case study involved the at-line monitoring of an HT micro-bioreactor system cultivating two mammalian cell cultures expressing two different therapeutic proteins. The spectra generated were analysed using a partial least squares (PLS) model. This enabled the successful prediction of the glucose, lactate, antibody, and viable cell density concentrations directly from the Raman spectra without reliance on multiple off-line analytical devices and using only a single low-volume sample (50–300 μL). However, upon the subsequent investigation of these models, only the glucose and lactate models appeared to be robust based upon their model coefficients containing the expected Raman vibrational signatures. On the downstream front, the HT Raman device was incorporated into the development of a cation exchange chromatography step for an Fc-fusion protein to compare different elution conditions. PLS models were derived from the spectra and were found to predict accurately monomer purity and concentration. The low molecular weight (LMW) and high molecular weight (HMW) species concentrations were found to be too low to be predicted accurately by the Raman device. However, the method enabled the classification of samples based on protein concentration and monomer purity, allowing a prioritisation and reduction in samples analysed using A280 UV absorbance and high-performance liquid chromatography (HPLC). The flexibility and highly configurable nature of this HT Raman spectroscopy microscope makes it an ideal tool for bioprocess research and development, and is a cost-effective solution based on its ability to support a large range of unit operations in both upstream and downstream process operations.

【 授权许可】

Unknown