Method Development for Multiplexing CGE-LIF Based Glycoanalysis

Analysis of Virus-Host Cell Interaction on the Proteome Level

Motivation

This project was related to other genomic and metabolic approaches, all focusing on the dynamic interaction of viruses with their mammalian host cells during vaccine production. Therefore, changes in the expression of host cell proteins in various mammalian cells after infection with different virus strains were characterized. In addition, proteome changes related to cultivation conditions were investigated.

Figure 1: Process scheme for differential protein abundance analysis and protein identification, using the 2D-DIGE system and nanoHPLC-MS/MS.

Aim of the project

Using two dimensional differential gel electrophoresis and mass spectrometry (Figure 1) we identified differentially expressed host cell proteins over the course of an infection phase. Different cell lines, like Vero and MDCK cells, are hosts for cell-based vaccine production. The results were correlated with integrated process data and gave detailed insight into vaccine production on the cellular level. This knowledge contributed to a better characterization und understanding of the complexity of virus-host cell interactions.

Publications

Vester, D.; Rapp, E.; Gade, D.; Genzel, Y.; Reichl, U.: Quantitative analysis of cellular proteome alterations in human influenza A virus-infected mammalian cell lines. Proteomics 9 (12), pp. 3316 - 3327 (2009)
Vester, D.; Rapp, E.; Kluge, S.; Genzel, Y.; Reichl, U.: Virus-host cell interactions in vaccine production cell lines infected with different human influenza A virus variants: A proteomic approach. Journal of Proteomics 73 (9), pp. 1656 - 1669 (2010)
Kluge, S.; Rourou, S.; Vester, D.; Majoul, S.; Benndorf, D.; Genzel, Y.; Rapp, E.; Kallel, H.; Reichl, U.: Proteome analysis of virus-host cell interaction: rabies virus replication in Vero cells in two different media. Applied Microbiology and Biotechnology 97 (12), pp. 5493 - 5506 (2013)
Kluge, S.; Genzel, Y.; Laus, K.; Serve, A.; Pflugmacher, A.; Peschel, B.; Rapp, E.; Reichl, U.: Ezrin and HNRNP expression correlate with increased virus release rate and early onset of virus-induced apoptosis of MDCK suspension cells. Biotechnology Journal 11 (10), pp. 1332 - 1342 (2016)
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