Adeno associated virus (AAV) capsids are a leading modality for in vivo gene delivery. Complete and precise characterization of capsid particles, including capsid and vector genome concentration, is necessary to safely and efficaciously dose patients. In virus development and production, it is important to determine the virus concentration at different stages of the process, to optimize the clone used as well as the production yields. Biolayer Interferometry (BLI) using AAV specific biosensors is a rapid label-free method for quantitation of AAV serotypes.
Gator AAVX Probes
The Gator AAVX probes are a high specificity antibody-based biosensors that enable direct capture and quantitation of different serotypes of AAV in crude lysates, column eluates, cell lysates and cell culture supernatants, serving as an alternative to traditional time-consuming analytical methods, such as qPCR, ddPCR, Dot blot and ELISA. The Gator AAVX probe uses proven CaptureSelect™ (Thermo Fisher Scientific) high affinity and high specificity anti-AAVX antibody.
- Dynamic range: 1 x 109–1 x 1013 vp/mL for most AAV serotypes
- Throughput: 8 samples in 10 minutes, 96 samples in 120 minutes
- Limit of detection: typically, 5 x 108 vp/mL (serotype dependent)
- Crude sample tolerant
- Stable over broad pH range
- Cost effective – Reusable at least 10 times by regeneration
The AAV2 serotype range was tested using the Gator AAVX probes. The data shows 4 orders of magnitude dynamic range. The data was acquired using 1:3 dilution in 10 min at 1,000rpm.
Accuracy and reproducibility
AAV5 serotype was used to test the accuracy and reproducibility. The table below shows recovery close to 100% and CV ranging from around 1.5% at medium and high titers to 10.9% at low titer.
Binding rate of different AAV serotypes
The binding rate of 8 different serotypes of AAV were studied. The figure below shows binding rates of tested serotypes at 2 x 1011 vp/mL concentration.
Data below shows performance after 10 regenerations of the same probe. No loss in binding rate observed even after 10 regenerations
Gator Bio Total Solution
GatorTM AAVX Probes
GatorTM Prime OR GatorTM Plus
Gator Bio Software
PL168-160017: GatorTM AAVX Probe