Comprehensive AAV characterisation using orthogonal methodologies for empty/full capsid analysis and genome integrity

ASGCT 2024 -- Adeno-associated virus (AAV) remains the viral vector of choice for in vivo gene therapy. However, there continue to be safety concerns regarding AAV therapeutics. The empty:full ratio of AAV vectors has been highlighted as an important critical quality attribute (CQA). It is also important to characterise the integrity and purity of the viral genome to ensure the safety and efficacy of the product. In this study, we acquired commercial research-grade AAV2-eGFP samples; two lots of 'full' capsids and one lot of 'empty' capsids. We assessed a portfolio of analytical methodologies and technologies to assess the empty:full capsid ratio as well as the integrity and purity of the viral genome. A summary of the analysis of other CQAs are also presented but are not the focus of the discussion.