de.mpg.escidoc.pubman.appbase.FacesBean
English
 
Help Guide Disclaimer Contact us Login
  Advanced SearchBrowse

Item

ITEM ACTIONSEXPORT

Released

Poster

Influence of Upstream Process Conditions on Influenza A Virus Hemagglutinin N-Glycosylation Pattern during Vaccine Production

MPS-Authors
http://pubman.mpdl.mpg.de/cone/persons/resource/persons86475

Schwarzer,  J.
Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons86442

Rapp,  E.
Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society;

http://pubman.mpdl.mpg.de/cone/persons/resource/persons86448

Reichl,  U.
Otto-von-Guericke-Universität Magdeburg;
Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society;

Locator
There are no locators available
Fulltext (public)
There are no public fulltexts available
Supplementary Material (public)
There is no public supplementary material available
Citation

Schwarzer, J., Rapp, E., & Reichl, U. (2008). Influence of Upstream Process Conditions on Influenza A Virus Hemagglutinin N-Glycosylation Pattern during Vaccine Production. Poster presented at 4th Glycan Forum, Berlin, Germany.


Cite as: http://hdl.handle.net/11858/00-001M-0000-0013-9567-7
Abstract
The envelope of influenza A virus is spiked with two glycoproteins: hemagglutinin (HA) and neuraminidase (NA). HA as the most abundant protein on the virus surface, triggers the strongest immune response. Each HA monomer contains 3 to 9 N-linked glycans, depending on the virus strain. The functional role of these glycans is still not completely understood. However, previous glycosylation studies have shown that structural modifications of these glycans can influence virus attachment to the host cell, and therefore change viral replication dynamics and its immunogenicity. The glycosylation pattern of viral proteins is affected by the glycosylation machinery of the host cell and their cultivation conditions. Further modifications in the structure can occur during inactivation and downstream processing steps. Hence, monitoring influenza virus HA glycosylation pattern in vaccine production processes can be used e.g. as a tool to ensure the immunogenicity of the antigens. In this study, a sensitive and reproducible N-glycan profiling method for cell culture derived influenza virus HA is presented. We are able to monitor potential variations of the glycosylation pattern, concerning N-glycan type and amount, due to the production process. The method includes virus purification directly from cell culture supernatant, protein separation by SDS-PAGE, endo- and exoglycosidase-cleavage of N-glycans, desalting and capillary gel electrophoresis with laser induced fluorescence detection (CGE-LIF). HA N-glycosylation is analyzed on two levels: first generating fingerprints and second performing structural analysis by spiking N-glycans with known structures as well as enzymatic sequencing. The impact of host cells used for virus production on HA N-glycosylation is presented. In addition, data from comparative analyses of HA N-glycan fingerprints of different influenza A virus strains and subtypes are shown. The developed method presents a promising tool to characterize and compare N-glycosylation patterns of HA during the major steps of up- and downstream processing in influenza virus vaccine production.