Viral Glycocalix in Dependence on Host Cell Line, Influenza Virus Strain and Production Process Parameters

Viral Glycocalix in Dependence on Host Cell Line, Influenza Virus Strain and Production Process Parameters


Glycoproteins, such as viral membrane protein hemagglutinin (HA) - the main antigen of influenza vaccines - can be considered as a collection of different glycoforms varying in structure of attached sugar residues (microheterogeneity) as well as in glycosylation site occupancy (macroheterogeneity). Since it is known that the glycosylation may significantly impact protein characteristics such as protein stability, antigenicity, immunogenicity, activity regulatory agencies usually demand glycosylation profiles for biopharmaceutical quality assurance. Since so far, there is only little known about the impact of glycosylation on vaccine efficiency and safety such regulations not yet exist for vaccines. However, first studies indicate a significant impact of glycosylation on vaccine immunogenicity, too.

Aim of the project

Using the developed high performance glycoanalysis system, the glycosylation patterns of viral surface proteins from unit operations in up- and downstream processing can be characterized. The main questions addressed regarding the glycosylation of HA, which is the most abundant and immunogenic influenza glycoprotein, are e.g.: the influence of host cell lines and virus strains [1,2], the impact of adaptation processes [3,4], changes due to modifications in cultivation conditions [5,6,7] and its impact on immunogenicity [8].


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Schwarzer, J.; Rapp, E.; Hennig, R.; Genzel, Y.; Jordan, I.; Sandig, V.; Reichl, U.: Glycan analysis in cell culture-based influenza vaccine production: Influence of host cell line and virus strain on the glycosylation pattern of viral hemagglutinin. Vaccine 27 (32), pp. 4325 - 4336 (2009)
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Rödig, J. V.; Rapp, E.; Bohne, J.; Kampe, M.; Kaffka, H.; Bock, A.; Genzel, Y.; Reichl, U.: Impact of Cultivation Conditions on N-glycosylation of Influenza Virus A Hemagglutinin Produced in MDCK Cell Culture. Biotechnology and Bioengineering 110 (6), pp. 1691 - 1703 (2013)
Hütter, J.; Rödig, J. V.; Höper, D.; Seeberger, P. H.; Reichl, U.; Rapp, E.; Lepenies, B.: Toward Animal Cell Culture-based Influenza Vaccine Design: Viral Hemagglutinin N-Glycosylation Markedly Impacts Immunogenicity. The Journal of Immunology 190 (1), pp. 220 - 230 (2013)
Hennig, R.; Rapp, E.; Kottler, R.; Cajic, S.; Reichl, U.: N -Glycosylation Fingerprinting of Viral Glycoproteins by xCGE-LIF. In: Carbohydrate-Based Vaccines: Methods and Protocols, pp. 123 - 143. Springer, New York (2015)

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