The One-Bac 4.0-derived AAVs present low levels of impurities and are free of rcAAV. 

Data from second-generation sequencing (illumina) and third-generation sequencing (PacBio) confrm that Genevoyager's One-Bac 4.0-derived AAVs exhibit high purity. 

DNA was extracted from the culture supernatant after three rounds of cell culture,and rcAAV was detected by qPCR.The results demonstrated the following:

For the positive control samples(wtAAV2/9)at 100 IU and 10IU,Ct values progressively decreased over the three amplification rounds,indicating successful amplification of rcAAV during culture.

For rAAV2/9 produced using the One-Bac 4.0 system,with inoculation doses of 1E+11vg and 1E+10vg,no significant decrease in Ct values was observed after three rounds.The Ct values remained comparable to the negative control(NC),indicating no detectable rcAAV.

The One-Bac 4.0-derived AAVs maintain high yields both before and after purification at a production scale of 50L.

The One-Bac 4.0 system achieves an impressive full capsid ratio of over 90% in its final product.

Results from cell and animal experiments demonstrate that One-Bac 4.0-derived AAV exhibits equivalent or higher infectivity compared to HEK293 AAV.

The Baculovirus Expression Vector (BEV) employed in the One-Bac 4.0 system maintains high stability over multiple passages and during long-term storage at ≤-80℃ for over 24 months,ensuring consistent high yields and robust expression levels of AAV batches at commercial scales.

With a consistent recovery rate of ≥30% from 5L to 200L scale-up, One-Bac 4.0 system demonstrates good scalability and effciency for large-scale production.

AAVs produced using the One-Bac 4.0 system exhibit low aggregation even at high concentrations. Stability assessments,including ddPCR-GOl and main peak SEC-HPLC,demonstrate that product quality remains stable under 4℃ and withstands multiple freeze-thaw cycles.

The One-Bac 4.0 system offers a more cost-effective approach to AAV production by replacing costly GMP plasmids and transfection reagents with a streamlined process for baculovirus vector production, significantly reducing the cost of materials for AAV vector manufacturing.

The establishment of a virus seed bank facilitates a rapid and simplified process for viral infection during production. This feature becomes more prominent as the scale of virus production increases, resulting in reduced timelines for production and delivery.