Adeno-associated viruses (AAVs) have emerged as promising vectors for gene therapy due to their low immunogenicity and ability to transduce a variety of cell types [1]. However, the development of safe and effective AAV-based therapeutics necessitates accurate and comprehensive characterization of these vectors. Key challenges in AAV analysis include determining particle size distribution, distinguishing between full and empty capsids, accurately measuring vector titer, and identifying protein impurities [1] [2]. Traditional methods often fall short in providing the required resolution and accuracy [3]. This application note presents an advanced analytical approach using Multi-Detector Asymmetrical Flow Field-Flow Fractionation (MD-AF4) to address these challenges. MD-AF4 offers a gentle, high-resolution, label-free technique for detailed evaluation of AAV8 vectors. The findings demonstrate the effectiveness of MD-AF4 in providing robust, reliable results that align with expectations and manufacturing requirements, offering a powerful tool for AAV characterization in gene therapy research and development.
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