Membrane Chromatography-Based Downstream Processing for Cell-Culture Produced Influenza Vaccines

Abstract

New influenza strains are constantly emerging, causing seasonal epidemics and raising concerns to the risk of a new global pandemic. Since vaccination is an effective method to prevent the spread of the disease and reduce its severity, the development of robust bioprocesses for producing pandemic influenza vaccines is exceptionally important. Herein, a membrane chromatography-based downstream processing platform with a demonstrated industrial application potential was established. Cell culture-derived influenza virus H1N1/A/PR/8/34 was harvested from benchtop bioreactor cultures. For the clarification of the cell culture broth, a depth filtration was selected as an alternative to centrifugation. After inactivation, an anion exchange chromatography membrane was used for viral capture and further processing. Additionally, two pandemic influenza virus strains, the H7N9 subtype of the A/Anhui/1/2013 and H3N2/A/Hong Kong/8/64, were successfully processed through similar downstream process steps establishing optimized process parameters. Overall, 41.3–62.5% viral recovery was achieved, with the removal of 86.3–96.5% host cell DNA and 95.5–99.7% of proteins. The proposed membrane chromatography purification is a scalable and generic method for the processing of different influenza strains and is a promising alternative to the current industrial purification of influenza vaccines based on ultracentrifugation methodologies.

Intensified Influenza Virus Production in Suspension HEK293SF Cell Cultures Operated in Fed-Batch or Perfusion with Continuous Harvest

“…Therefore, the development of robust downstream processes is critical to support cell culture-based influenza vaccines. A robust and scalable process for the purification of influenza viruses produced in HEK293SF cells is described [ 49 ], reporting up to 96.5% and 99.7% removal of host cell DNA and proteins, respectively.…”

Getting Hold of the Tobamovirus Particle – Why and How? Purification Routes over Time and a New Customizable Approach

“…Last but not least, despite most frequent applications in final polishing steps with discardible devices, membrane chromatography is increasingly valued for virus manufacturing due to its scalability, and may also be used in different flow-through modes depending on the type of device [310]. Membrane chromatography, compared to traditional resin column chromatography, reduces pressure drop and mass transfer resistance [311,312]. This is similar to the advantages of the interconnected channels in monolithic columns, which -apart from uses in bind/elute chromatography (see 2│5│2) -may also be integrated into advanced radial flow devices for upscaling, thereby evading problems with the mechanical instability of disk-shaped monolithic CIM media or pressure increase in elongated columns [310].…”

Getting Hold of the Tobamovirus Particle—Why and How? Purification Routes over Time and a New Customizable Approach

“…Finally, despite the most frequent applications in the final purification steps involving discardable devices, membrane chromatography is increasingly valued for virus manufacturing due to its scalability and may also be used in different flow-through modes depending on the type of device [ 322 ]. Membrane chromatography, compared to traditional resin column chromatography, reduces pressure drop and mass transfer resistance [ 323 , 324 ]. This is similar to the advantages of the interconnected channels in monolithic columns, which—apart from uses in bind/elute chromatography (see Section 2.5.2 )—may also be integrated into advanced radial flow devices for upscaling, thereby evading problems with the mechanical instability of disk-shaped monolithic CIM media or pressure increase in elongated columns [ 322 ].…”