Description
Biopharmaceutical manufacturing processes often release host cell proteins (HCP) during upstream processing, that may affect the safety, quality, stability and efficacy of the final product. Hence, regulatory authorities require extensive monitoring of HCPs during downstream protein purification. However, current detection methods such as enzyme-linked immunosorbent assay have several limitations due to differences in quantity and affinity of anti-HCP polyclonal antibodies for various HCP species. Therefore, it is crucial to develop orthogonal methods for quantitative HCP analysis.Here, we provide a bottom-up proteomics workflow for quantitative HCP analysis, employing comprehensive strong anion exchange, reversed phase liquid chromatography tandem mass spectrometry. This study has important implications for understanding the relationship between the LC efficacy, the peptide quantification and bioprocess-related HCP removal strategies, ultimately leading to increased protein identification. For that, the resolving power of each LC dimension was optimized for model peptides towards maximized peak capacity per unit time, whilst maintaining short total analysis times.
Ongoing studies will provide further insight into the impact of enhanced LC performance on monitoring the clearance potential of high risk, or low-intensity HCPs by data independent acquisition mass spectrometry. As proof of concept, we demonstrate HCP identification in bioprocess-related samples of various complexities, provided by different host cell lines.
The ultimate objective of our study is to identify and monitor the clearance of HCPs in a novel DSP workflow for sIgA. Quantitative and qualitative data on these HCPs will offer further inside, that can guide the process development towards targeted removal strategies and overall process efficiency.
| Periode | 6 nov 2023 |
|---|---|
| Evenementstitel | INTERNATIONAL SYMPOSIUM ON THE SEPARATION OF PROTEINS, PEPTIDES AND POLYNUCLEOTIDES |
| Evenementstype | Conference |
| Conferentienummer | 42 |
| Locatie | Vienna, Austria |
| Mate van erkenning | International |