AN2004: Why and how to quantify AAV aggregates by FFF-MALS

The aggregate content is a critical quality attribute (CQA) of AAV-based gene therapies. This application note compares analytical separations by size exclusion chromatography (SEC) and field-flow fractionation (FFF), along with detection by UV, fluorescence, and multi-angle light scattering (MALS). The results demonstrate that FFF-MALS is the most appropriate method for quantifying aggregates of AAV-mediated products, from dimer, trimer, and small oligomers to large aggregates.