A flow-through chromatography process for influenza A and B virus purification

Area of Science:

• Biotechnology
• Virology
• Downstream Processing

Background:

• Influenza virus infections pose a significant public health threat, necessitating effective vaccines.
• Cell culture-based production of influenza vaccines requires efficient purification methods.
• Existing purification processes can be costly and sensitive to virus strains.

Purpose of the Study:

• To develop an economic, robust, and efficient downstream process for influenza virus purification.
• To establish a chromatography-based method that avoids virus strain-specific capture steps.
• To meet the stringent purity requirements for human vaccine production.

Main Methods:

• A three-step chromatographic process was developed: anion exchange chromatography (AEC), Benzonase(®) treatment, and ligand-activated core (LCC) size exclusion chromatography.
• The process was tested using two influenza A virus strains and one influenza B virus strain.
• Purification efficiency was evaluated based on virus yield, protein contamination, and DNA depletion.

Main Results:

• The process achieved high influenza virus yields (≥68%) for all tested strains.
• Protein contamination levels met the European Pharmacopeia requirements for vaccine production.
• DNA was depleted by ≥98.7%, with concentrations near the 10ng/dose limit.
• Added Benzonase(®) was successfully removed from the final product.

Conclusions:

• The developed downstream process is a simple, robust, and economic platform for cell culture-derived influenza vaccine production.
• This method offers a viable alternative to existing purification strategies, ensuring high purity and yield.
• The process is adaptable for various influenza virus strains, supporting vaccine manufacturing.