Two questions, what do you consider typical titer for AAV production and what can you do to increase it?
This question is part of the following Ask The Expert session:
Ask the Expert – Transfection Best Practices for AAV Gene Therapy
Job Title: Applications Manager
Typically, AAV production titers are around 10e11 to 10e12 in VG/ml and 10e8 to 10e9 TU/ml. To put these numbers into perspective, it’s important to keep in mind that two AAV serotypes will most likely not give the same yield, even when using the same production process. AAV production yields vary depending on the serotype and the gene of interest. The best way to increase production of a given AAV is to go over critical parameters that directly have an impact on the yield: plasmid DNA, transfection reagent, cells and medium.
- Plasmid DNA– using a DOE approach, transgene/packaging/helper plasmid ratios, as well as the final DNA amount per million cells can be refined to improve yield.
- Transfection reagent – comparing transfection reagents to reach highest production yield, and further refining the plasmid DNA/ transfection reagent ratio to improve delivery efficiency and gene expression.
- Cells and medium-choosing the best cell culture system for highest production yield for the scale that you aim for, adherent or suspension-based.