Comparison of Wildtype and Recombinant Adeno-Associated Virus (AAV) Production Reveals Distinct Productivity and Content Ratio Profiles.
- 1Department of Chemical and Biomolecular Engineering, University of Delaware, Newark, Delaware, USA.
Abstract
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 × 10¹² vector genomes/mL culture and 72.2% full, compared to recombinant transfection systems (1.63 × 10¹¹ vector genomes/mL and 26.9% full).
Related
- Streamlined Production of Recombinant Adeno-Associated Viruses Using Gateway Cloning Technology for Neural Circuit All-Optical Interrogation
- A biosensor toolbox for monitoring recombinant protein production in Escherichia coli.
- Complement receptor C3aR marks heterogeneous tumor‐associated macrophage states associated with improved survival in IDH ‐wildtype glioblastoma
- Estimation of agricultural carbon abatement costs using a marginal-productivity-based production-input model: evidence from Fujian Province, China
- Recombinant bacterial β-galactosidases for lactose bioconversion and galacto-oligosaccharide production: a comparative overview
BibTeX
@article{alfieri2026comparison,
title = {{Comparison of Wildtype and Recombinant Adeno-Associated Virus (AAV) Production Reveals Distinct Productivity and Content Ratio Profiles.}},
author = {Sofia Alfieri and Kelvin H Lee},
journal = {Biotechnology journal},
year = {2026},
month = oct,
doi = {10.1002/biot.70331},
url = {https://europepmc.org/article/MED/42961188},
note = {CC BY-NC-ND},
}
RIS
TY - JOUR
TI - Comparison of Wildtype and Recombinant Adeno-Associated Virus (AAV) Production Reveals Distinct Productivity and Content Ratio Profiles.
AU - Sofia Alfieri
AU - Kelvin H Lee
PY - 2026
DA - 2026/10/30
JO - Biotechnology journal
DO - 10.1002/biot.70331
UR - https://europepmc.org/article/MED/42961188
AB - 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 × 10¹² vector genomes/mL culture and 72.2% full, compared to recombinant transfection systems (1.63 × 10¹¹ vector genomes/mL and 26.9% full).
SN - 1860-6768
SN - 1860-7314
ER -