The Avitide AVIPure® Residual Ligand Assay kits are intended for use in quantitating the Avitide AVIPure®-AAV8 Ligand. This kit (Item Number F1005) incorporates a well-qualified sample treatment method to dissociate the ligand from the AAV virus.
AVIPure-AAV8 Ligand, immobilized on various chromatography media, is used to purify AAV8 virus. Even when covalently attached, the ligand can leach off of the chromatography support and co-elute with AAV8. Residual AVIPure-AAV8 Ligand must be minimized to avoid any adverse patient effects.
The Avitide AVIPure® — AAV8 Ligand assay is a two-site immuno-enzymatic assay. Samples containing AVIPure® — AAV8 Ligand are mixed with sample denaturing buffer to dissociate the ligand from the AAV virus. The samples are then reacted simultaneously with a horseradish peroxidase (HRP) enzyme labeled anti-AVIPure® antibody in microtiter strips coated with an affinity purified capture antibody. The immunological reactions result in the formation of a sandwich complex of solid phase antibody-Ligand-enzyme labeled antibody. After a wash step to remove any unbound reactants, the strips are then reacted with tetramethylbenzidine (TMB) substrate. The amount of hydrolyzed substrate is read on a microtiter plate reader and will be directly proportional to the concentration of AVIPure® — AAV8 Ligand present in the sample. Accurate quantitation is achieved by comparing the signal of unknowns to AVIPure® — AAV8 Ligand standards assayed at the same time.
This immunoassay provides a simple to use, precise, and highly sensitive method to detect AVIPure-AAV8 Ligand to less than 1ng/mL. As such, it can be used as a tool to aid in optimal purification process development and in routine quality control of in-process streams as well as final product.
Please contact Repligen to inquire about AVIPure® – AAV8 Resin.