We are excited to announce that our first AAVone paper has been accepted for publication in MTNA(Molecular Therapy – Nucleic Acids)!
The AAVone system consolidates the adenoviral helper genes, AAV packaging genes, and the vector transgene cassette into a single, compact plasmid with a 13-kb pAAVone backbone. Compared to the traditional triple-plasmid system, AAVone streamlines the production process by eliminating the need for fine-tuning plasmid ratios. It delivers a 2- to 4-fold increase in AAV yield, ensures low batch-to-batch variability, and reduces DNA input requirements while maintaining favorable full capsid ratios and minimizing impurities. Importantly, the AAVone system does not introduce a risk of generating rcAAV.
At the upcoming hashtag#ASGCT meeting, our paper’s first author, Dr. Rongze Yang, will present additional data on the scalability and Process Optimization of the AAVone platform at poster #1974.
The AAVone system can be efficiently scaled from small flasks to pilot and large-scale bioreactors(XDR200) without compromising productivity. With further process optimization—including improved HEK 293 cell lines, advanced transfection reagents and enhancers, and optimized culture media—AAVone has achieved yields of up to 5 × 10¹⁵ vector genomes per liter (VG/L) for AAV9 production. Under these conditions, the AAV yield per cell approaches 1 × 10⁶ vector genomes, given a cell density of approximately 5 × 10⁶ cells/mL at the end point.
These findings raise a fundamental question: What ultimately limits AAV productivity? After extensive optimization efforts, scientists have achieved production titers of up to 1 × 10¹⁵ VG/L using triple-plasmid transfection methods. With our AAVone system, we are now able to reach 5 × 10¹⁵ VG/L.
We believe a critical factor in maximizing yield is ensuring that all essential genetic components are delivered into the same cell. The AAVone system addresses this challenge by consolidating all necessary elements into a single plasmid. This design creates a theoretical linear correlation between transfection efficiency and AAV yield. In contrast, the conventional triple-plasmid system is inherently less efficient—only cells that receive all three plasmids can produce AAV vectors. This results in a nonlinear and less predictable relationship between transfection efficiency and overall vector output.
By continuing to optimize the individual components involved in AAV production, we believe it’s possible to surpass 1 × 10¹⁶ VG/L, pushing the boundaries of current manufacturing capability.
In summary, the AAVone platform is a highly efficient, scalable, cost-effective, and consistent solution for AAV vector production, making it particularly well-suited for GMP-grade applications.
We welcome inquiries from those interested in testing and licensing this novel technology.