Quantification of influenza A virus proteins during replication in MDCK cells
Motivation
Madin-Darby canine kidney (MDCK) cells are widely used in basic research and for production of cell culture-derived influenza vaccines. To identify targets for antiviral therapies and to optimize vaccine manufacturing, a detailed understanding of the viral life cycle is important. Here, we focus on propagation of influenza A virus (IAV). This includes the characterization of the virus entry, the synthesis of the various viral RNAs and proteins, the transfer of viral compounds in the cell and virus budding. Our experimental studies are complemented with mathematical modeling approaches. However, while quantitative data is available for virus entry as well as viral mRNA, vRNA and cRNA accumulation in infected cells, such data is mostly lacking for the synthesis of viral proteins and their accumulation on the intra- and extracellular level.
Aim of the project
While further developing existing models for IAV propagation [1,2], we acquire detailed quantitative information about IAV protein expression using state-of-the-art mass spectrometry methods (see Figure below). Employing influenza A/PR/8/34 (H1N1) as a model strain, we track the most abundant IAV proteins during infection and investigate virus-host cell dynamics for a range of IAV strains and host cells considered for vaccine production.