Adenovirus 5 recovery using nanofiber ion-exchange adsorbents

Area of Science:

• Biotechnology
• Materials Science
• Chemical Engineering

Background:

• Viral vectors are crucial for vaccines and gene therapy, but their high-quality manufacturing is challenging.
• Current therapeutic protein purification methods using chromatography are effective but require adaptation for viral vectors due to size and stability differences.

Purpose of the Study:

• To characterize a novel cellulose nanofiber ion-exchange sorbent for viral vector separation.
• To investigate the impact of quaternary amine ligand density on viral recovery, stability, and separation resolution.
• To establish a basis for efficient viral vector purification using optimized chromatographic conditions.

Main Methods:

• Utilized cellulose nanofiber ion-exchange sorbents derivatized with quaternary amine ligands at varying densities (440–1029 µmol/g).
• Assessed the effect of ligand density and adsorption duration (24 min vs. 1 min) on infective viral product recovery and stability.
• Evaluated the impact of ligand density and elution strategy on the separation resolution, focusing on the virus particle to infectious virus particle ratio.

Main Results:

• Higher ligand densities reduced viral product stability, especially with prolonged contact times; this was mitigated by high flow rates (1 min cycle time).
• Ligand density and elution strategy significantly influenced the virus particle to infectious virus particle ratio, a key indicator of viral quality.
• The developed sorbent demonstrated potential for separating viral vectors, impurities, and process-related components.

Conclusions:

• Cellulose nanofiber ion-exchange chromatography offers a promising approach for viral vector purification.
• Optimizing sorbent surface chemistry (ligand density) and process parameters (flow rate, elution) is critical for maximizing viral recovery and quality.
• This method provides a foundation for scalable and efficient purification of labile viral vectors used in gene therapy and vaccine development.