title: Mono- and polyprotic buffer systems in anion exchange chromatography of influenza virus particles.
abstract: Different ions typically used in downstream processing of biologicals are evaluated for their potential in anion exchange chromatography of an industrially produced, pandemic influenza H1N1 virus. Capacity, selectivity and recovery are investigated based on single step elution parallel chromatography experiments. The inactivated H1N1 feedstream is produced in Madin-Darby Bovine Kidney cells. Interesting effects are found for sodium phosphate and sodium citrate. Both anions are triprotic kosmotropes. Anion exchange chromatography generally offers high scalability to satisfy sudden demands for vaccines, which may occur in case of an emerging influenza outbreak. Appropriate pH conditions for H1N1 adsorption are determined by Zeta potential measurements. The dynamic binding capacity of a salt tolerant polyamine-type resin is up to 6.4 times greater than the capacity of a grafted Q-type resin. Pseudo-affinity interactions of polyamines with the M2 protein of influenza may contribute to the obtained capacity increase. Both resins achieve greater capacity in sodium phosphate buffer compared to Tris/HCl. A recovery of 67% and DNA clearance close to 100% without DNAse treatment are achieved for the Q-type resin. Recovery of the virus from the salt tolerant resin requires the use of polyprotic acids in the elution buffer. 85% of the DNA and 60% of the proteins can be removed by the salt tolerant resin. The presence of sodium phosphate during anion exchange chromatography seems to support stability of the H1N1 particles in presence of hydrophobic cations.
authors.affiliation: Tosoh Bioscience GmbH, Im Leuschnerpark 4, 64347 Griesheim, Germany. Electronic address: Judith.Vajda@tosoh.com.
authors.forename: Judith
authors.initials: J
authors.lastname: Vajda
authors.affiliation: Karlsruher Institut für Technologie, Institut für Bio- und Lebensmitteltechnik, Engler-Bunte Ring 3, 76131 Karlsruhe, Germany. Electronic address: dennis.weber2@student.kit.edu.
authors.forename: Dennis
authors.initials: D
authors.lastname: Weber
authors.affiliation: IDT Biologika GmbH, Am Pharmapark, 06861 Dessau-Roßlau, Germany. Electronic address: Sabine.Stefaniak@idt-biologika.de.
authors.forename: Sabine
authors.initials: S
authors.lastname: Stefaniak
authors.affiliation: IDT Biologika GmbH, Am Pharmapark, 06861 Dessau-Roßlau, Germany. Electronic address: Boris.Hundt@idt.biologika.de.
authors.forename: Boris
authors.initials: B
authors.lastname: Hundt
authors.affiliation: University of Applied Sciences Mannheim, Institut für Biochemie, Paul-Wittsack-Str. 10, 68163 Mannheim, Germany. Electronic address: TanjaRathfelder@gmx.de.
authors.forename: Tanja
authors.initials: T
authors.lastname: Rathfelder
authors.affiliation: Tosoh Bioscience GmbH, Im Leuschnerpark 4, 64347 Griesheim, Germany. Electronic address: Egbert.mueller@tosoh.com.
authors.forename: Egbert
authors.initials: E
authors.lastname: Müller
mesh_terms: D000327:Adsorption; D000818:Animals; D000838:Anions; D002021:Buffers; D002417:Cattle; D002852:Chromatography, Ion Exchange / Q000379:methods; D006863:Hydrogen-Ion Concentration; D053118:Influenza A Virus, H1N1 Subtype / Q000302:isolation & purification*; D007252:Influenza Vaccines; D012965:Sodium Chloride; D014763:Viral Matrix Proteins / Q000737:chemistry; D014771:Virion / Q000302:isolation & purification*; D000086082:Viroporin Proteins
chemical_list: D000838:Anions; D002021:Buffers; D007252:Influenza Vaccines; D012965:Sodium Chloride; D014763:Viral Matrix Proteins; C488937:M2 protein, Influenza A virus; D000086082:Viroporin Proteins
keywords: Anion exchange chromatography; Multimodal chromatography; Pandemic influenza virus; Polyprotic acids; Pseudo affinity chromatography; Whole virus particle capturing