HOST CELL PROTEIN CHARACTERISATION AND MASS SPECTROMETRY STRATEGIES

Описание: Presented by Xuezhi Bi, Group Leader (Proteomics) /Senior Scientist at Bioprocessing Technology Institute, A*STAR

Host cell proteins (HCPs) are process-related contaminants that are released to the cell culture supernatant due to the secretion or lysis of host cells during the production of biologics. Due to the potential adverse effects of HCPs on the biotherapeutics stability, quality and efficacy, monitoring and clearance of HCPs is essential in manufacturing processes. Multi-analyte ELISA is the ‘gold standard’ method routinely used to detect and measure the amount of residual HCP impurities in the biologics. However, multi-analyte ELISA is expensive, laborious, and unable to detect weak and non-immunoreactive HCPs as well as to provide identity and quantity of individual HCP species. Thus, there is an increasing demand for orthogonal analytical tools, such as 2-D western blot and LC-MS/MS, to identify and quantify HCPs in the in-process intermediates and final products. Among these techniques, LC-MS/MS provides a high throughput, robust and sensitive platform to identify and quantify individual HCP, including those of low abundance. The incorporation of a quantitative MS/MS workflow supported by a comprehensive spectral library in bioprocessing strategies would enable real-time monitoring during upstream production and downstream purification of biologics, which would provide valuable information to the biopharmaceutical industry to produce safe and efficacious drugs.