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Application: AAV with AAV2 ITR qPCR titration
Background: Recombinant adeno-associated virus (rAAV) is a widely used vector in gene therapy applications. rAAVs exist in the nucleus as episomes and are unable to integrate into the cellular genome. While this brings safety benefits, as there is no risk of random integration and activation of oncogenes, the cell division rate dictates the timeframe for transgene loss. Their application is thus particularly useful for the delivery of transgenes to post-mitotic tissues like the brain, retinal, liver, skeletal muscle, and the heart. The payload carried by AAVs is somewhat smaller than other viral tools use for transduction, usually smaller than 5 kb. AAV titers can be measured as a physical titer, infectious titer, or ratio of full to empty capsids. Accurately determining the genome titer is essential not only for setting clinical doses but also as a foundational step in various analytical assays for characterizing AAV products. While AAV genome titers, physical titers, are typically measured by qPCR (quantitative polymerase chain reaction), precision has remained a challenge despite numerous refinements to the process. Typically, standard curves are done with plasmid DNA, which does not provide a direct correlation with material of interest. The use of an assay that relies on digestion-free method, includes an AAV standard and reference material, is well-suited for adoption in gene therapy research and can be further automated for high-throughput applications.
Description: The AAV qPCR Titration Kit is a kit designed to measure AAV (adeno-associated virus) titers of particles with AAV2 ITR (inverted terminal repeats) by qPCR (quantitative polymerase chain-reaction). It offers a streamlined, one-step process without needing digestion steps. Engineered for high sensitivity and specificity, it minimizes non-specific background, enhancing performance over comparable kits. The kit contains enough reagents for 100 reactions. It includes a separate ROX (carboxyrhodamine), a passive fluorescence reference dye that allows for signal normalization, and an AAV reference material as qPCR standard for precise quantification.This product has been validated across multiple AAV samples with varying serotypes, purification status, and encapsulated transgenes, consistently demonstrating high levels of accuracy, precision, and robustness.Need custom AAV design and production? Visit our AAV service page for more information.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed. Components are shipped in dry ice and stored at -20°C freezer for long term storage. The components maintain their stability and performance after 5 freeze-thaw cycles.
Target: AAV
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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