Improving AAV and LV upstream manufacturing using cGMP lipid polymer nanocomplexes (LPNCs)
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With success of cell and gene therapies and exponentially increasing demand, the second wave of viral vector manufacturing is focused on large-scale suspension cell culture and/or adherent cell culture within intensified fixed bed bioreactors. In response to this need, Mirus Bio developed a novel HEK 293 cGMP cell transfection technology that increases AAV and LV titers 2-10-fold over existing technologies. The transfection reagent formulation is a mixture of lipid and polymer which enables the formation of lipid polymer nanocomplexes (LPNCs), for high efficiency transfection of multiple plasmids as well as higher recombinant AAV and LV titers. Complementary enhancers and/or complex formation solutions that excel in suspension cultures further augment AAV or LV yields.
Important characteristics were considered during reagent and enhancer development to enable scalability, straightforward optimization and seamless adoption within multiple manufacturing AAV and LV workflows. The VirusGEN® system has ample complex formation time, with no loss in titer out to 60 minutes. Due to the targeting specificity of the AAV capsid protein and its influence on cell tropism, multiple AAV serotypes were tested for both titer and virus quality. As with AAV workflows, high titer lentivirus production can be obtained across different cell types and serum free complete media formulations with TransIT-VirusGEN® Reagent and the VirusGEN® LV Kit. Scalability is also possible with the VirusGEN® in stirred tank bioreactors.
Data will be presented that demonstrate that VirusGEN® pushes the limits of high titer recombinant AAV and LV production to provide more usable virus per run, maximizing precious time and resources.
- Learn about a new cGMP transfection platform option in the cell and gene therapy space for viral vector development.
- How this unique cGMP transfection platform:
- Is compatible across multiple media and cell types
- Yields higher functional LV titers over existing cGMP competitors on the market
- Generates both higher functional titers and % full capsid across multiple serotypes
- How improvements in the transient transfection step of viral vector production runs can significantly decrease cost per dose
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