Nanopore Sequencing Grants Detailed Insights Into A ​Small Molecule's Impact On Encapsidated DNA​ Composition During Manufacturing Platform Development

By K. Breunig, M. Haubner, F. Dunker-Seidler, F. Sonntag, E. Schweigert, A. Schulze, and M. Hoerer

A key challenge in recombinant adeno-associated virus (rAAV) gene therapy is the cost-efficient production of large quantities of high-quality vectors. Ascend identified a small molecule (SM-016) capable of increasing vector titers, but its impact on vector quality required further investigation. Efficient characterization of rAAV vector batches is essential for platform and process development, and nanopore sequencing offers a fast, comprehensive method with low input requirements.

This study presents data on vector length distribution, encapsidated DNA impurities, and vector integrity. Results show that SM-016 does not negatively affect vector length distribution, integrity, or host cell DNA impurities. However, a slight increase in plasmid backbone-derived impurities was observed, attributed to elevated ITR read-through/bidirectional packaging, suggesting areas for optimization.

These findings emphasize the value of nanopore sequencing in refining vector production platforms.