Improved adeno-associated virus empty and full capsid separation using weak partitioning multi-column AEX...

Area of Science:

• Biotechnology
• Bioprocessing
• Chromatography

Background:

• Separating empty and full recombinant adeno-associated virus (rAAV) capsids is critical for gene therapy.
• Anion exchange chromatography (AEX) is a key purification step, but requires optimization for efficiency.

Purpose of the Study:

• To develop and evaluate a new AEX method combining weak partitioning chromatography (WPC) and multi-column chromatography (MCC) for improved rAAV purification.
• To enhance the percentage of full rAAV capsids and overall productivity in the AEX step.

Main Methods:

• Implementation of a novel AEX method integrating WPC technology for selective capsid displacement during loading.
• Utilization of MCC for parallel column processing to increase throughput.
• Comparison of the AEX-WPC-MCC method against baseline AEX batch chromatography.

Main Results:

• The AEX-WPC-MCC method achieved approximately a 20% increase in the full rAAV percentage in the AEX pool.
• A similar increase of approximately 20% was observed in rAAV genome recovery.
• Productivity of the AEX step increased by approximately 34-fold compared to the baseline batch method.

Conclusions:

• The AEX-WPC-MCC method offers significant improvements in full rAAV capsid separation and purification efficiency.
• This integrated chromatography approach has the potential to enhance large-scale rAAV manufacturing.
• The method may contribute to reducing the cost of goods for rAAV-based gene therapies.