Comparison of Wildtype and Recombinant Adeno-Associated Virus (AAV) Production Reveals Distinct Productivity and Content Ratio Profiles

Recombinant adeno-associated virus is a leading viral vector for gene therapy. However, recombinant adeno-associated virus production systems have low titer and content ratio (ratio of genome-containing to total adeno-associated virus capsids), making it difficult to generate the large number of vector genomes required for clinical or commercial use. In contrast, wildtype adeno-associated virus achieves over ten-fold higher titer compared to recombinant adeno-associated virus and a content ratio of over 95% full. Despite the higher productivity of wildtype adeno-associated virus, the mechanisms behind its productivity remain poorly understood. In this study, we developed and characterized a novel wildtype adeno-associated virus production platform in suspension human embryonic kidney cells across varying multiplicities of infection of adeno-associated virus to adenovirus. The impact of cellular viral genome delivery mechanisms and the interplay between adeno-associated virus and its adenoviral helper were investigated by comparing infection-based systems with transfection-based and infection-transfection hybrid systems. Wildtype and hybrid infection systems had remarkably different host cell growth profiles and viral gene expression patterns, and achieved over a log-fold higher titers and 45% higher content ratios, reaching 8.1 × 10 12 vector genomes/mL culture and 72.2% full, compared to recombinant transfection systems (1.63 × 10 11 vector genomes/mL and 26.9% full).

Abstract from PubMed (PMID 42961188). This entry was added automatically by our daily literature monitor because it matches the topics we follow; read the full paper at the original source.