Optimization and Scale up Strategies for Reproducible AAV Enrichment Step on CIMmultus QA HR Line

Rok Žigon, Sara Drmota Prebil, Tomaž Švigelj, Kaja Kozjak, Timotej Žvanut, Maja Leskovec, Aleš Štrancar

Gene Therapy, 2026

Abstract:
AAV gene therapy advancement encounters challenges in separating empty, full, and intermediate capsids and scaling downstream processing. This study highlights monolithic chromatography’s role in maintaining high flow rates and resolution, making it suitable for large-scale applications. CIMmultus® HR columns provide high reproducibility across purification scales, achieving ± 3% isoconductivity elution of empty AAV2/8 capsids. The improved method enriches full capsids to 88% ± 9% with 77% ± 3% genome recovery. CIM® QA HR Monolithic Well Plate screening and single-column experiments validate these findings, supporting step elution approaches. The optimized AAV polishing method successfully scales from 1 to 8000 mL using CIM® products, ensuring consistent recovery and purity.