Improving AAV8 purification with continuous affinity capture: From batch to continuous multicolumn chromatography.

Area of Science:

• Biotechnology
• Chemical Engineering
• Molecular Biology

Background:

• Gene therapy relies heavily on adeno-associated viruses (AAVs), necessitating efficient and scalable purification methods.
• Current purification processes face challenges in meeting the increasing demand for high-quality AAVs.

Purpose of the Study:

• To evaluate and compare batch multicolumn chromatography (B-MCC) and continuous sequential multicolumn chromatography (S-MCC) for adeno-associated virus type 8 (AAV8) purification.
• To optimize AAV purification using the Resolute® BioSMB PD platform for enhanced recovery, productivity, and resource utilization.

Main Methods:

• Exploration of breakthrough behavior to define a continuous loading strategy for chromatography.
• Development and modification of batch multicolumn chromatography (B-MCC) and continuous sequential multicolumn chromatography (S-MCC) processes.
• Comparison of B-MCC and S-MCC based on recovery, productivity, buffer consumption, and resin utilization.

Main Results:

• The S-MCC process achieved a higher AAV recovery of 76% at the highest evaluated load concentration.
• S-MCC demonstrated 1.5 times higher productivity (1.8 × 10^17 VP h^-1 mL^-1) compared to B-MCC.
• S-MCC halved buffer consumption relative to B-MCC for equivalent processed feed volumes.

Conclusions:

• Continuous sequential multicolumn chromatography (S-MCC) significantly improves AAV purification efficiency, offering superior recovery and productivity.
• S-MCC presents a more resource-efficient purification strategy, reducing buffer consumption and enhancing resin utilization.
• These findings support the adoption of continuous processing for high-demand biologics like AAVs in the biopharmaceutical industry.