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Despite the differences in influenza virus surface antigens and virulence of seasonal subtypes, manufacturers are well\u2010adapted to respond to this periodical vaccine demand. Due to decades of influenza virus research, the development of new influenza vaccines is relatively straight forward. In similarity with the ongoing coronavirus disease 2019 pandemic, vaccine manufacturing is a major bottleneck for a rapid supply of the billions of doses required worldwide. In particular, egg\u2010based vaccine production would be difficult to schedule and shortages of other egg\u2010based vaccines with high demands also have to be anticipated. Cell culture\u2010based production systems enable the manufacturing of large amounts of vaccines within a short time frame and expand significantly our options to respond to pandemics and emerging viral diseases. In this study, we present an integrated process for the production of inactivated influenza A virus vaccines based on a Madin\u2013Darby Canine Kidney (MDCK) suspension cell line cultivated in a chemically defined medium. Very high titers of 3.6 log<jats:sub>10<\/jats:sub>(HAU\/100\u2009\u00b5l) were achieved using fast\u2010growing MDCK cells at concentrations up to 9.5\u2009\u00d7\u200910<jats:sup>6<\/jats:sup> cells\/ml infected with influenza A\/PR\/8\/34 H1N1 virus in 1\u2009L stirred tank bioreactors. A combination of membrane\u2010based steric\u2010exclusion chromatography followed by pseudo\u2010affinity chromatography with a sulfated cellulose membrane adsorber enabled full recovery for the virus capture step and up to 80% recovery for the virus polishing step. Purified virus particles showed a homogenous size distribution with a mean diameter of 80\u2009nm. Based on a monovalent dose of 15\u2009\u00b5g hemagglutinin (single\u2010radial immunodiffusion assay), the level of total protein and host cell DNA was 58\u2009\u00b5g and 10\u2009ng, respectively. Furthermore, all process steps can be fully scaled up to industrial quantities for commercial manufacturing of either seasonal or pandemic influenza virus vaccines. Fast production of up to 300 vaccine doses per liter within 4\u20135 days makes this process competitive not only to other cell\u2010based processes but to egg\u2010based processes as well.<\/jats:p>","DOI":"10.1002\/bit.27876","type":"journal-article","created":{"date-parts":[[2021,7,5]],"date-time":"2021-07-05T04:15:09Z","timestamp":1625458509000},"page":"3996-4013","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":20,"title":["Towards integrated production of an influenza A vaccine candidate with MDCK suspension cells"],"prefix":"10.1002","volume":"118","author":[{"ORCID":"https:\/\/orcid.org\/0000-0002-6901-6161","authenticated-orcid":false,"given":"Thomas","family":"Bissinger","sequence":"first","affiliation":[{"name":"Bioprocess Engineering Max Planck Institute for Dynamics of Complex Technical Systems Magdeburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3616-3026","authenticated-orcid":false,"given":"Yixiao","family":"Wu","sequence":"additional","affiliation":[{"name":"Bioprocess Engineering Max Planck Institute for Dynamics of Complex Technical Systems Magdeburg Germany"},{"name":"State Key Laboratory of Bioreactor Engineering East China University of Science and Technology Shanghai China"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-5540-4454","authenticated-orcid":false,"given":"Pavel","family":"Marichal\u2010Gallardo","sequence":"additional","affiliation":[{"name":"Bioprocess Engineering Max Planck Institute for Dynamics of Complex Technical Systems Magdeburg Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dietmar","family":"Riedel","sequence":"additional","affiliation":[{"name":"Facility for Transmission Electron Microscopy Max Planck Institute for Biophysical Chemistry Goettingen Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Xuping","family":"Liu","sequence":"additional","affiliation":[{"name":"Shanghai BioEngine Sci\u2010Tech Co. 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