Objective:

To present the development and characterization of the NBX1P01 clonal HEK293 cell line for improved recombinant adeno-associated virus (rAAV) production.

Key Findings:

• NBX1P01 produced a two-fold higher full-to-empty rAAV capsid ratio compared to a leading commercial HEK293 cell line.
• The clone remained genomically stable over 55 population doubling levels with lower genetic heterogeneity.
• Long-read sequencing confirmed over 70% intact rAAV genomes with minimal contaminating DNA.
• Consistent performance was demonstrated across production volumes from 10 mL to 1.5 L and supported multiple rAAV serotypes.

Interpretation:

The NBX1P01 cell line represents a significant advancement in rAAV manufacturing, addressing key challenges in scalability and product quality for gene therapy applications.

Limitations:

• The study does not address long-term performance beyond 55 population doublings.
• Further validation in diverse production settings may be required.

Conclusion:

NBX1P01 offers a reliable and consistent platform for high-quality rAAV production, essential for advancing gene therapy accessibility.