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|Product Name||AAV-8 Titration ELISA|
|Description||Enzyme Immunoassay for the Quantitative Determination of AAV Serotype 8 Particles in Cell Culture Supernatants and Purified Virus Preparations|
|Background||Adeno-associated virus (AAV) is a non pathogenic ssDNA virus that is a topic of intense study in gene therapy. The virus transduces a wide variety of dividing and non-dividing cells showing long-term gene expression with no cellular immune response. AAV has been used in several clinical trials (e.g. FIX, CFTR, Parkinson’s disease, Canavan disease) showing no serious vector-related adverse effects. Methods for the characterisation of AAV preparations currently include titration ELISA, realtime PCR, DNA dot blot, determination of transducing units, infectious center assay, SDSPAGE or electron microscopy.
Immunotitration by ARP's AAV8 Titration ELISA offers a fast, sensitive and reproducible method for titration of intact AAV8 wt virions, AAV8 recombinant virions or assembled and intact empty AAV8 capsids.
|Principle||The assay is based on the sandwich ELISA technique. A monoclonal antibody specific for a conformational epitope on assembled AAV8 capsids (ADK8) is coated onto microtiter strips and is used to capture AAV8 particles from the specimen. Captured AAV particles are detected in two steps. First a biotin-conjugated monoclonal antibody to AAV8 (ADK8) is bound to the immune complex. In the second step streptavidin peroxidase conjugate reacts with the biotin molecules. Addition of substrate solution results in a color reaction which is proportional to the amount of specifically bound viral particles. The absorbance is measured photometrically at 450 nm.
The kit control provided contains an AAV 2/8 particle preparation, including empty capsids. It shows a typical titration curve when used in dilutions of steps of two (Fig. 1). It allows the quantitative determination of samples of an unknown particle titer (immunological titer) and the calibration of an in-house AAV8 preparation (e.g. infectious titer, transducing units, DNA dot blot titer).
|Intended Use||For Research Use Only|
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