Figure 1.

Presentation of the AAV8/Anc82 Experimental Design Used to Study Capsid Mosaic Formation and Cross-Packaging

(A) Sequence variability represented at the surface of the AAV8 capsid (PDB: 2QA0). Variable residues between AAV8 and Anc82 are represented in red. (B) AAV8 and Anc82 VP1 sequence alignment between amino acid positions 586 and 591. AAV8 capsid-specific ADK8 antibodies are known to bind this motif, disrupted in the case of Anc82. (C) Thermal stability profiles of AAV8 (blue), Anc82 (orange), and a mix of both particles at a 1:1 vp ratio (gray, dashed line). Left: normalized fluorescence signals; right: derivative fluorescence signals. (D) Overview on the experimental procedure. AAV8/Anc82 libraries were produced through cotransfection of HEK293 cells with a decreasing amount of rep2/cap8 and rep2/capAnc82 plasmids, using either the pSL or pSub201 production system. AAVs were further harvested and subjected to a battery of assays to investigate cross-packaging and capsid mosaic formation.