A Scalable Analytical Method For Quality-Driven AAV Characterization

As adeno-associated virus (AAV) gene therapies advance toward larger patient populations and commercial manufacturing, accurate characterization of viral vector quality has become increasingly important. A key challenge is measuring capsid composition, as AAV preparations contain mixtures of full, empty, and partially filled capsids that can affect potency, dosing, safety, and regulatory acceptance. Among available analytical methods, anion exchange chromatography-high performance liquid chromatography (AEX-HPLC) has emerged as a practical, GMP-compatible approach for routine capsid analysis, offering high reproducibility, scalability, throughput, and compatibility with standard chromatography platforms.

AEX-HPLC enables simultaneous measurement of multiple critical quality attributes, including empty-to-full capsid ratios, genome-containing capsid fractions, and total capsid titer in a single analytical run. Beyond quantification, chromatographic profiles can provide insight into packaging efficiency, process consistency, and product heterogeneity, making the technique valuable throughout process development, purification optimization, technology transfer, and manufacturing scale-up. When used alongside orthogonal analytical methods such as analytical ultracentrifugation and charge detection mass spectrometry, AEX-HPLC provides robust, quantitative data that support regulatory expectations, lot release, comparability studies, and long-term process control, helping developers maintain consistent AAV product quality from early development through commercial production.