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  Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A viruses

Opitz, L., Salaklang, J., Büttner, H., Reichl, U., & Wolff, M. W. (2007). Lectin-affinity chromatography for downstream processing of MDCK cell culture derived human influenza A viruses. Vaccine, 25(5), 939-947. doi:10.1016/j.vaccine.2006.08.043.

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Item Permalink: http://hdl.handle.net/11858/00-001M-0000-0013-984F-9 Version Permalink: http://hdl.handle.net/11858/00-001M-0000-0014-A1F5-8
Genre: Journal Article

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Opitz, L.1, Author              
Salaklang, J.1, Author              
Büttner, H.2, Author
Reichl, U.1, 3, Author              
Wolff, M. W.1, Author              
Affiliations:
1Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, Max Planck Society, ou_1738140              
2Univ. of Applied Sciences, Muenster, Germany, persistent:22              
3Otto-von-Guericke-Universität Magdeburg, ou_1738156              

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 Abstract: The presented study aims on the development of a capture step for the purification of cell culture derived influenza viruses using lectin affinity chromatography. Human influenza A/Puerto Rico/8/34 virus produced in Madin Darby canine kidney cells have been chosen as a model. The influenza A virus envelop possesses two viral glycoproteins: hemagglutinin and neuraminidase. Oligosaccharides of theses glycoproteins can be targeted as affinity ligands using specific lectins. First, lectins have been screened via lectin blots and spin columns. Adequate lectins have been chosen based on published glycan structures of hemagglutinin. The most specific binding was achieved via the galactose specific Erythrina cristagalli and Euonymus europaeus lectins. Second, the chromatographic separations characteristics of these lectins have been further determined via FPLC. These experiments revealed that the rate of hemagglutinin glycan binding to the ligands was higher with the Euonymus europaeus compared to the Erythrina cristagalli lectin. Third, viral recoveries in addition to the total protein and contaminating host cell DNA have been balanced in a series of Euonymus europaeus lectin chromatography runs. The total protein and dsDNA content in the product fraction of the affinity chromatography was reduced from the starting conditions to 21% and 0.1%, respectively. The average viral recovery in the product fraction was 97%. SDS-PAGE analysis indicated that the majority of the eluted proteins were of viral origin. The reproducibility and column stability was confirmed in more than 25 runs applying 6 different virus product batches. © 2006 Elsevier Ltd. All rights reserved [accessed 2013 November 14th]

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Language(s): eng - English
 Dates: 2007
 Publication Status: Published in print
 Pages: -
 Publishing info: -
 Table of Contents: -
 Rev. Method: Peer
 Identifiers: eDoc: 285555
DOI: 10.1016/j.vaccine.2006.08.043
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Title: Vaccine
Source Genre: Journal
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Publ. Info: Guildford, Surrey, UK : Elsevier
Pages: - Volume / Issue: 25 (5) Sequence Number: - Start / End Page: 939 - 947 Identifier: ISSN: 0264-410X
CoNE: /journals/resource/954925498091