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AAV-8 Titration ELISA

AAV-8 Titration ELISA


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Product Code: PRAAV8


Product Details Protocol
Product Code: PRAAV8
Product: 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).
Storage: 2-8C
Intended Use: For Research Use Only
This product is sold for laboratory research use or further manufacturing only and should not be used for human therapeutic or diagnostic applications. The information presented is believed to be accurate; however, said information and products are offered without warranty or guarantee since the ultimate conditions of use and the variability of the materials treated are beyond our control. Nothing disclosed herein is to be construed as a recommendation to use our products in violation of any patents. Under no circumstances shall ARP American Research Products, Inc. be liable for damages, whether consequential, compensatory, incidental or special, strict liability or negligence, breach of warranty or any other theory arising out of the use of the products available from ARP American Research Products, Inc. Nothing contained herein warrants that the use of the products will not infringe on the claims of any patents covering the product itself or the use thereof in combination with other products or in the operation of any process.

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