Characterizing the Content and Structure of AAV Capsids by Size Exclusion Chromatography and Orbitrap-Based Charge Detection-Mass Spectrometry.

ABSTRACT: Adeno-associated virus (AAV) is currently the most widely used vector in gene therapy applications. However, a significant challenge in the manufacturing process of recombinant AAV (rAAV) is the presence of empty capsids, oligomers, aggregates, and partially filled capsids. These components do not provide any therapeutic benefit but add to the overall viral load, which could increase immunogenicity and reduce transduction efficiency. Here, we present a strategy that utilizes size exclusion chromatography (SEC) equipped with multiangle light scattering (MALS) and a diode-array detector (DAD), followed by orbitrap-based charge detection-mass spectrometry (CD-MS). The SEC step was used to separate AAV capsids (non-aggregates) from oligomers, aggregates, and low molecular weight contaminants.

SUBMITTER: Pathak K 

PROVIDER: S-EPMC12333375 | biostudies-literature | 2025 Aug

REPOSITORIES: biostudies-literature