Density gradient ultracentrifugation (DGUC) is a well-established method for separating empty (E) and full (F) AAV capsids based on density differences. However, traditional DGUC is labour-intensive and lacks direct detection capabilities, requiring manual fraction collection and subsequent analysis. These limitations can be overcome by integrating post-DGUC profiling with the PATfix® chromatography system. Compared to analytical ultracentrifugation (AUC), this approach offers a cost-effective and accessible alternative, eliminating the need for an expensive analytical ultracentrifuge while leveraging standard laboratory equipment. Additionally, it simplifies analysis by bypassing the complex data deconvolution and sedimentation velocity calculations required in AUC, instead relying on a stratigraphic readout from the centrifuge tube while maintaining high accuracy.
