title: Production of high-titer human influenza A virus with adherent and suspension MDCK cells cultured in a single-use hollow fiber bioreactor.
abstract: Hollow fiber bioreactors (HFBRs) have been widely described as capable of supporting the production of highly concentrated monoclonal antibodies and recombinant proteins. Only recently HFBRs have been proposed as new single-use platforms for production of high-titer influenza A virus. These bioreactors contain multiple hollow fiber capillary tubes that separate the bioreactor in an intra- and an extra-capillary space. Cells are usually cultured in the extra-capillary space and can grow to a very high cell concentration. This work describes the evaluation of the single-use hollow fiber bioreactor PRIMER HF (Biovest International Inc., USA) for production of influenza A virus. The process was setup, characterized and optimized by running a total of 15 cultivations. The HFBRs were seeded with either adherent or suspension MDCK cells, and infected with influenza virus A/PR/8/34 (H1N1), and the pandemic strain A/Mexico/4108/2009 (H1N1). High HA titers and TCID₅₀ of up to 3.87 log₁₀(HA units/100 μL) and 1.8 × 10(10)virions/mL, respectively, were obtained for A/PR/8/34 influenza strain. Influenza virus was collected by performing multiple harvests of the extra-capillary space during a virus production time of up to 12 days. Cell-specific virus yields between 2,000 and 8,000 virions/cell were estimated for adherent MDCK cells, and between 11,000 and 19,000 virions/cell for suspension MDCK.SUS2 cells. These results do not only coincide with the cell-specific virus yields obtained with cultivations in stirred tank bioreactors and other high cell density systems, but also demonstrate that HFBRs are promising and competitive single-use platforms that can be considered for commercial production of influenza virus.
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany; Advanced Materials and Processes (MAP), Elite Graduate Program, University of Erlangen-Nuremberg, Martenstr. 5-7, 91058 Erlangen, Germany. Electronic address: tapia@mpi-magdeburg.mpg.de.
authors.forename: Felipe
authors.initials: F
authors.lastname: Tapia
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany; University of Applied Sciences Esslingen, Kanalstrasse 33, 73728 Esslingen, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Vogel
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany.
authors.forename: Yvonne
authors.initials: Y
authors.lastname: Genzel
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany.
authors.forename: Ilona
authors.initials: I
authors.lastname: Behrendt
authors.affiliation: Biovest International Inc., 8500 Evergreen Blvd., 55433 Minneapolis, MN, USA.
authors.forename: Mark
authors.initials: M
authors.lastname: Hirschel
authors.affiliation: Biovest International Inc., 8500 Evergreen Blvd., 55433 Minneapolis, MN, USA.
authors.forename: J David
authors.initials: JD
authors.lastname: Gangemi
authors.affiliation: Max Planck Institute for Dynamics of Complex Technical Systems, Sandtorstr. 1, 39106 Magdeburg, Germany; Chair for Bioprocess Engineering, Otto-von-Guericke-University Magdeburg, Universitätsplatz 2, 39106 Magdeburg, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Reichl
mesh_terms: D000818:Animals; D019149:Bioreactors; D004285:Dogs; D053118:Influenza A Virus, H1N1 Subtype / Q000254:growth & development*; D061985:Madin Darby Canine Kidney Cells; D014776:Virus Cultivation / Q000379:methods*
keywords: Adherent cells; Hollow fiber bioreactor; Influenza; MDCK; MDCK.SUS2; Suspension cells