AAV6 K531 serves a dual function in selective receptor and antibody ADK6 recognition

Adeno-associated viruses (AAVs) are being developed as vectors for the treatment of genetic disorders. However, pre-existing antibodies present a significant limitation to achieving optimal efficacy for the AAV gene delivery system. Efforts aimed at engineering vectors with the ability to evade the immune response include identification of residues on the virus capsid important for these interactions and changing them. Here K531 is identified as the determinant of monoclonal antibody ADK6 recognition by AAV6, and not the closely related AAV1. The AAV6-ADK6 complex structure was determined by cryo-electron microscopy and the footprint confirmed by cell-based assays. The ADK6 footprint overlaps previously identified AAV antigenic regions and neutralizes by blocking essential cell surface glycan attachment sites. This study thus expands the available repertoire of AAV-antibody information that can guide the design of host immune escaping AAV vectors able to maintain capsid functionality. •ReciprocalAAV1 and AAV6 variants are comparable to the WT vectors in assembly, genome titer, and transduction efficiency.•The ADK6 footprint on the AAV6 capsid identified a single residue, K531,as the determinant of recognition.•ADK6 binding overlaps previously defined AAV epitopes and is predicted to sterically hinder glycan receptor interaction.