Rapid Quality Control Assessment of Adeno-Associated Virus Vectors Via Stunner

Abstract

As recombinant adeno-associated virus (AAV) vectors expand the availability of gene therapy; high-speed low-volume analytical methods become more vital for manufacturing. Current approaches for characterization are time consuming and sample consuming. Stunner combines ultraviolet–visible (UV–Vis) spectroscopy, static light scattering (SLS), and dynamic light scattering (DLS) to characterize up to 96 samples in <1 h, using 2 μL per sample. These data can be used to precisely quantify critical AAV titer, empty/full ratio, size, and aggregation. This study compares this technique with quantitative real-time PCR (qPCR), enzyme-linked immunoassay (ELISA), transmission electron microscopy, and analytical ultracentrifugation using AAV vectors with multiple serotypes, genome sizes, and purification strategies. Recommended sample criteria were established to provide guidelines on the suitability of Stunner analysis on a per-sample basis. The data presented herein lay the groundwork in establishing the use of combined UV–Vis, SLS, and DLS as a powerful and flexible analytical tool for AAV gene therapy vector development and manufacturing.