Proteomics in Downstream Processing of Influenza Virus
An increasing number of biopharmaceuticals are manufactured by recombinant technologies using a specific host-cell system. A critical part of the development of a marketed biopharmaceutical is the analysis of impurities and degradation products. Generally, the composition of biopharmaceuticals – except formulation components – is classified into product-related substances, product-related impurities, and process-related impurities .
Although some impurities are benign in terms of toxicity, others may be allergenic. Residual host-cell proteins (HCPs) are a major component of process-related impurities that have been reported to cause immune responses in patients at levels as low as 10-100 ppm . Unlike any other kind of therapeutic drugs, vaccines are administered to healthy individuals, making the safety assessment of this family of products of great significance.
Determining the identities and contents of HCPs is critical for downstream processes. However, since they are specific to each manufacturing workflow, their analysis is complicated and usually requires multiple analytical methodologies that are based on chromatography, spectroscopy, and Mass Spectrometry (MS). The latter can provide the most definitive identifications in either purified, enriched or complex protein mixtures. MS-based proteomics can be divided into bottom-up, shotgun, and top-down approaches.
Aim of the project
The aim of this project is the identification of proteinaceous impurities during downstream processing of cell culture-derived influenza virus particles with an Orbitrap mass spectrometer for the purpose of toxicity/safety assessment of vaccines.
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