Enhanced Downstream Processing for a Cell-Based Avian Influenza (H5N1) Vaccine

Area of Science:

• Virology
• Biotechnology
• Vaccine Development

Background:

• H5N1 highly pathogenic avian influenza virus (HPAIV) poses a significant global health threat.
• Existing H5N1 HPAIV vaccines face limited approval due to safety concerns, particularly MDCK cell tumorigenicity.
• Minimizing residual DNA and host cell proteins (HCP) is crucial for enhancing vaccine safety.

Purpose of the Study:

• To develop an improved downstream processing method for cell-based H5N1 HPAIV vaccines.
• To enhance vaccine safety by significantly reducing residual DNA and HCP levels.
• To ensure the integrity and efficacy of the H5N1 HPAIV antigen.

Main Methods:

• A two-step chromatographic process was developed for vaccine purification.
• Capto™ Core 700, a multimodal resin, was utilized for polishing.
• Hydrophobic-interaction chromatography (HIC) with polypropylene glycol facilitated virus particle capture.

Main Results:

• Virus recovery achieved 68.16% following the two-step chromatographic process.
• Host cell protein (HCP) levels were reduced to 2112.60 ng/mL.
• Residual DNA levels were reduced to 6.4 ng/mL.
• Western blot, HPLC, and TEM confirmed antigen presence, spherical shape, and appropriate particle size.

Conclusions:

• The developed two-step downstream process is effective in reducing DNA and HCP in cell-based H5N1 HPAIV vaccines.
• This method ensures the quality and safety of the H5N1 HPAIV antigen.
• The process shows potential as an efficient and cost-effective platform for cell-based influenza vaccines.