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One-Bac 4.0 system is an insect/baculovirus system for AAV production. The production process consists of several key parts: baculovirus plasmid construction, seed virus production, seed virus amplification, seed virus infection of insect cells, and AAV harvest. This system achieves linear amplification of production by infecting insect cells with baculovirus containing the AAV genome, overcoming the bottleneck of large-scale AAV preparation. One-Bac 4.0 system has the potential to benefit more patients due to its high yield compared to other AAV production systems. 

One-Bac 4.0 System New Patent Group

One-Bac 4.0 system process flow chart


QC index for AAV

Genevoyager is equipped with a one-stop gene therapy CDMO platform, alongsidean experienced GMP quality control teamwithexpertise inmethodology development, validation/verification, and testing toensure the high quality of our products.The routine quality release tests and methods are outlined below.


Category

Test Item

Test Method

Identification and structural analysis

DNA identification

Sanger sequencing, Next-generation sequencing (NGS), Third-generation sequencing (TGS)

Capsid protein identification

Western blot, SDS-PAGE, CE, LC-MS

Biological activity

Potency

In vitro cell-based biological activity, Animal studies

Content

Virus capsid titer

ELISA

Genome titer

ddPCR, qPCR

Infectious titer

TCID50

VG:IU

Ratio of genome titer to infectioustiter

Purity

Total protein

microBCA

Capsid protein purity

SDS-PAGE, CE

Viral monomer purity

SEC-HPLC

Empty capsid ratio

AUC, TEM, AEX-HPLC, SEC-MALS

Particle size

DLS

Impurities

Residual Sf9 host cell DNA

qPCR

Residual Sf9 host cell protein

ELISA

Residualnuclease

ELISA

Residualinfectious baculovirus

Plaque assay, TCID50

Residualbaculovirus DNA

qPCR

Residualrhabdovirus

qPCR

Specific process-related residues (affinity ligand, Triton, Tween20, etc.)

ELISA, UV, HPLC

Safety

Replication-competent AAV (rcAAV)

Culturemethod+ qPCR

Mycoplasma

qPCR, Culture method

Spiroplasma

qPCR, Culture method

Endotoxin

Gel electrophoresis, Spectrophotometry

Sterility

Membrane filtration, Direct inoculation

Abnormal toxicity

Injectionin mouse and guinea pig 




More Content of Interest

High Yields:

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


High Passage Stability of BEV:

The Baculovirus Expression Vector (BEV) employed in the One-Bac 4.0 system maintains high stability for multiple passages, ensuring consistent high yield and infectious activity of AAV batches during large-scale production.


High Full Capsid Ratio:

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


High Infectious Activity:

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