| dc.description.abstract | This project explored a variable pathlength UV absorbance-based Process Analytical
Technology tool (SoloVPE®) for quantifying Vector Genome titer and estimating the
percentage of full capsids in AAV samples, aiming to enhance Critical Quality Attributes
monitoring in Adeno-Associated Virus production. Results from SoloVPE® were
compared to an established orthogonal method, Size Exclusion Chromatography
coupled to a Multi-Angle Light Scattering detector (SEC-MALS) to evaluate accuracy and
reproducibility. Initial findings indicated that SoloVPE® results correlate well with SECMALS, maintaining a deviation within ±10%. However, performance was varied across
sample types and storage conditions. For VG quantification, SoloVPE® exhibited
excellent linearity (R² > 0.99) which was maintained over at least a one-log range of
concentrations, validating its use in dynamic manufacturing environments. On the other
hand, % full capsid estimations showed less reproducibility, also in comparison to the
orthogonal Mass Photometry platform, and a tendency for underestimation compared to
SEC-MALS results. The limitations of SoloVPE® stem from its inability to distinguish
between full and partially filled capsids, alongside potential stability issues related to
sample handling and storage. Despite these challenges, the SoloVPE® oƯers significant
advantages in speed and ease of use, positioning it as a valuable PAT tool for in-process
monitoring and screening in AAV production. | |
