title: A flow-through chromatography process for influenza A and B virus purification.
abstract: Vaccination is still the most efficient measure to protect against influenza virus infections. Besides the seasonal wave of influenza, pandemic outbreaks of bird or swine flu represent a high threat to human population. With the establishment of cell culture-based processes, there is a growing demand for robust, economic and efficient downstream processes for influenza virus purification. This study focused on the development of an economic flow-through chromatographic process avoiding virus strain sensitive capture steps. Therefore, a three-step process consisting of anion exchange chromatography (AEC), Benzonase(®) treatment, and size exclusion chromatography with a ligand-activated core (LCC) was established, and tested for purification of two influenza A virus strains and one influenza B virus strain. The process resulted in high virus yields (≥68%) with protein contamination levels fulfilling requirements of the European Pharmacopeia for production of influenza vaccines for human use. DNA was depleted by ≥98.7% for all strains. The measured DNA concentrations per dose were close to the required limits of 10ng DNA per dose set by the European Pharmacopeia. In addition, the added Benzonase(®) could be successfully removed from the product fraction. Overall, the presented downstream process could potentially represent a simple, robust and economic platform technology for production of cell culture-derived influenza vaccines.
authors.affiliation: Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany. Electronic address: weigel@mpi-magdeburg.mpg.de.
authors.forename: Thomas
authors.initials: T
authors.lastname: Weigel
authors.affiliation: Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany; Faculty of Pharmaceutical Biotechnology, Biberach University of Applied Sciences, 88400 Biberach, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Solomaier
authors.affiliation: Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany; Department of Biotechnology, Lausitz University of Applied Sciences, 01968 Senftenberg, Germany.
authors.forename: Alessa
authors.initials: A
authors.lastname: Peuker
authors.affiliation: Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany.
authors.forename: Trinath
authors.initials: T
authors.lastname: Pathapati
authors.affiliation: Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany; Chair of Bioprocess Engineering, Otto-von-Guericke University, 39106 Magdeburg, Germany.
authors.forename: Michael W
authors.initials: MW
authors.lastname: Wolff
authors.affiliation: Bioprocess Engineering, Max Planck Institute for Dynamics of Complex Technical Systems, 39106 Magdeburg, Germany; Chair of Bioprocess Engineering, Otto-von-Guericke University, 39106 Magdeburg, Germany.
authors.forename: Udo
authors.initials: U
authors.lastname: Reichl
mesh_terms: D000818:Animals; D002853:Chromatography, Liquid / Q000191:economics / Q000379:methods*; D006801:Humans; D009980:Influenza A virus / Q000302:isolation & purification*; D009981:Influenza B virus / Q000302:isolation & purification*; D007252:Influenza Vaccines / Q000302:isolation & purification*; D013678:Technology, Pharmaceutical / Q000191:economics / Q000379:methods*; D014773:Virology / Q000191:economics / Q000379:methods*
chemical_list: D007252:Influenza Vaccines
keywords: Downstream process; Flow-through chromatography; Influenza virus purification; Strain independency; Vaccine production