Binding Sites and Reactivity
For AAV9 the neutralization epitope has been identified and might give an additional indication about the binding residues of the neutralizing antibody ADK9 used for the corresponding AAV9 Titration ELISAs. ADK9 recognizes and neutralizes assembled AAV9 capsids. Is is unsure if ADK9 actually binds at aa453-457 but since no other epitope has been identified in epitopemapping, it is very likely.
The ADK9 and the ADK9-1R antibody were tested for neutralization using different antibody concentrations. After the HeLa cells were seeded in a 96-well plate they were incubated for 48h with the AAV9 particles containing a GFP reporter and the indicated concentration of the ADK9 or the ADK9-1R antibody, respectively.
The microscopic analysis of the cells clearly demonstrates the neutralizing activity of the ADK9 and ADK9-1R antibody by decreasing GFP expression with increasing antibody concentration.
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- Tseng, Y.S. et al. Generation and characterization of anti-Adeno-associated virus serotype 8 (AAV8) and anti-AAV9 monoclonal antibodies. J. Virol. Meth. 236, 105-110 (2016)
- Varadi, K. et al. Novel random peptide libraries displayed on AAV serotype 9 for selection of endothelial cell-directed gene transfer vectors. Gene Ther. 19, 800-9 (2012).
