AAVpro Concentrator
AAVpro Concentrator
Brand: Takara Bio.
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AAVpro Concentrator
Efficient concentration of AAV particles from cell culture supernatant
AAVpro Concentrator provides a simple, highly efficient alternative to ultracentrifugation for concentrating AAV particles of any serotype from cell culture supernatant. In contrast with other concentration methods that involve only precipitation of AAV particles, AAVpro Concentrator incorporates filtration steps to yield AAV preps of higher purity. The scalable mix-and-spin protocol can accommodate a range of input volumes and requires less than one hour of hands-on time. The AAVpro Concentrator reagent is provided as a 4X concentrate, which can be applied in proportion to the amount of cell culture supernatant being processed. The kit includes reagents and filtration columns for processing up to 150 ml of culture supernatant for two samples.
Overview
- Efficient concentration of AAV particles from cell culture supernatant
- Filtration steps included for removal of impurities
- Simple mix-and-spin protocol requires less than one hour of hands-on time
- Scalable method enables processing of various input volumes
- No ultracentrifugation required
- Compatible with any AAV serotype
- Can be used to process culture medium with or without serum
Applications
- Concentration of AAV from cell culture supernatant
- Transfer of AAV particles from one buffer to another
- Rescue of low-titer supernatants
- In vitro or in vivo transduction using AAV vectors
The AAVpro Concentrator process for AAV vector enrichment
An outline of the AAV vector enrichment process using AAVpro Concentrator.
Concentration and recovery rate of AAV1 vectors concentrated using AAVpro Concentrator
Concentration and recovery rate of AAV1 vectors, concentrated using AAVpro Concentrator. In this experiment, culture supernatant containing AAV1 vectors expressing ZsGreen1 was concentrated from a starting input of 40 ml (one T225 flask) down to a final volume of 200 μl using AAVpro Concentrator, for a 95-fold increase in AAV titer and a recovery rate of ~48%.
Biological titer of AAV vectors concentrated using AAVpro Concentrator, as determined by FACS analysis
AAV titration in transducing units, as determined by FACS analysis of ZsGreen1 expression in HT1080 cells. The cells were infected with AAV at 5,000 or 500 vg/cell (MOI) and analyzed by flow cytometry after three days. Concentrated AAV particles administered at MOIs of 5,000 or 500 were able to infect cells at rates of 44.7% or 9.2%, respectively.
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The AAVpro Concentrator process for AAV vector enrichment
An outline of the AAV vector enrichment process using AAVpro Concentrator.
Concentration and recovery rate of AAV1 vectors concentrated using AAVpro Concentrator
Concentration and recovery rate of AAV1 vectors, concentrated using AAVpro Concentrator. In this experiment, culture supernatant containing AAV1 vectors expressing ZsGreen1 was concentrated from a starting input of 40 ml (one T225 flask) down to a final volume of 200 μl using AAVpro Concentrator, for a 95-fold increase in AAV titer and a recovery rate of ~48%.
Biological titer of AAV vectors concentrated using AAVpro Concentrator, as determined by FACS analysis
AAV titration in transducing units, as determined by FACS analysis of ZsGreen1 expression in HT1080 cells. The cells were infected with AAV at 5,000 or 500 vg/cell (MOI) and analyzed by flow cytometry after three days. Concentrated AAV particles administered at MOIs of 5,000 or 500 were able to infect cells at rates of 44.7% or 9.2%, respectively.
