Probing recombinant AAV capsid integrity and genome release after thermal stress by mass photometry.

ABSTRACT: Adeno-associated viruses (AAVs) are gaining traction as delivery vehicles for gene therapy although the molecular understanding of AAV-transgene release is still limited. Typically, the process of viral uncoating is investigated (in vitro) through thermal stress, revealing capsid disintegration at elevated temperatures. To assess the (in)stability of different empty and filled AAV preparations, we used the light-scattering-based interferometric microscopy technique of mass photometry that, on a single-particle basis, determines the molecular weight of AAVs. By introducing a heat-stable DNA plasmid as an internal standard, we quantitatively probed the impact of heat on AAVs. Generally, empty AAVs exhibited greater heat resistance than genome-filled particles. Our data also indicate t

SUBMITTER: Ebberink EHTM 

PROVIDER: S-EPMC11295964 | biostudies-literature | 2024 Sep

REPOSITORIES: biostudies-literature