Abstract
Pharmaceutical proteins, compared to small molecular weight drugs, are relatively fragile molecules, thus necessitating monitoring protein unfolding and aggregation during production and post-marketing. Currently, many analytical techniques take offline measurements, which cannot directly assess protein folding during production and unfolding during processing and storage. In addition, several orthogonal techniques are needed during production and market surveillance. In this study, we introduce the use of time-gated Raman spectroscopy to identify molecular descriptors of protein unfolding. Raman spectroscopy can measure the unfolding of proteins in-line and in real-time without labels. Using K-means clustering and PCA analysis, we could correlate local unfolding events with traditional analytical methods. This is the first step toward predictive modeling of unfolding events of proteins during production and storage.
| Original language | English |
|---|---|
| Article number | 1639 |
| Number of pages | 18 |
| Journal | Pharmaceutics |
| Volume | 14 |
| Issue number | 8 |
| DOIs | |
| Publication status | Published - Aug 2022 |
| MoE publication type | A1 Journal article-refereed |
Funding
This research was funded by the Academy of Finland project 303884 and VTT technical research Centre of Finland. L.G. gratefully acknowledges the support of the Drug Discovery and Chemical Biology Network of Finland. J.I. has also been funded by grants from the Finnish Cultural Foundation, the Evald and Hilda Nissi Foundation, the Päivikki and Sakari Sohlberg Foundation, and the Paulo Foundation. D.P. received Erasmus+ program funding from the Italian Ministry of Education.
Keywords
- biologics
- CD
- DLS
- in-line measurement
- K-means clustering
- PCA
- pharmaceutical proteins
- protein unfolding
- Raman spectroscopy
- tryptophan fluorescence