|Targeted Proteomics of Influenza Viral Particles|
|Wed 11:30 AM - Session: Molecular Diagnostics|
|Christopher M. Colangelo|
|Determining the absolute protein levels of both virus-encoded and host-encoded proteins in influenza virions will provide crucial information on virus structure and the stoichiometry of the individual protein components, which may reveal new understanding of influenza biology. Influenza virions contain nine viral proteins; however, analyses of other enveloped viruses have revealed that proteins from the host cell can also be detected in virions.
Our previous work (Shaw et al. 2008), used a combination of MudPIT and Gel electrophoresis to identify the 9 essential virus-encoded proteins, as well as 36 host-encoded proteins in Influenza A/WSN/33 virus grown in vero kidney cells. The resulting peptide identification data was used to synthesize internal control peptides for all 9 influenza proteins and 6 of the 36 host encoded proteins. Applied Biosystems 4000 QTRAP triple quadrupole mass spectrometer and the MRMPilot software was then utilized to develop a Multiple Reaction Monitoring (MRM) method for determining the absolute levels of the virus-encoded influenza virion proteins and relative levels of host-encoded proteins.