Improving Viral Vector Concentration for Gene Therapy
Author Information
Author(s): Christine V Ichim, Richard A Wells
Primary Institution: University of Toronto
Hypothesis
Can successive rounds of ultracentrifugation increase the concentration of viral vectors for better transduction of difficult-to-infect cells?
Conclusion
The method of concentrating virus using multiple rounds of ultracentrifugation significantly increases viral titers, enabling effective transduction of bone marrow cells.
Supporting Evidence
- Four rounds of ultracentrifugation resulted in an additive increase in viral titer.
- VSV-G coated lentiviral particles maintained transduction efficiency after multiple rounds of concentration.
- Transduction rates in primary bone marrow cells improved significantly with concentrated viral stocks.
Takeaway
This study shows that by spinning virus samples multiple times in a centrifuge, we can make them stronger and better at getting into cells, which is important for gene therapy.
Methodology
The study involved generating stable retrovirus producer cell lines, producing large volumes of virus-containing supernatant, and testing transduction ability after multiple rounds of ultracentrifugation.
Limitations
The study did not establish the maximum achievable viral titers or the effects of prolonged centrifugation on viral stability.
Digital Object Identifier (DOI)
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