Speakers:
Andreja Gramc Livk, Head of Process Analytics, Sartorius BIA Separations
Sebastijan Peljhan, Head of Basic Research, Sartorius BIA Separations
Abstract:
This webinar will explore empty-to-full AAV capsid ratio determination using multimodal detection techniques in combination with improved anion exchange (AEX) chromatography methods and post-ultracentrifugation density gradient profiling.
First, participants will learn about an AEX analytical method involving different elution strategies for empty and full capsid separation based on modifying mobile phases with organic solvents and Mg organic salts. The method allows baseline separation of AAV capsid subspecies and enables characterization of empty, full, partially filled, and other capsid species in different sample matrices.
Second, we will explore the complementary capabilities of post-density gradient ultracentrifugation profiling for characterization of AAV materials. Compared to analytical ultracentrifugation (AUC), this technique offers a cheaper and more straightforward alternative, as it eliminates the need for an expensive analytical ultracentrifuge and can be performed on regular laboratory ultracentrifuge. Additionally, it avoids the complex data deconvolution and sedimentation velocity analysis required in AUC.
Data will be presented demonstrating how multimodal detection setups, including UV absorbance, light scattering, and fluorescence, deliver precise insights into the empty-to-full capsid ratio, capsid integrity, and particle concentration. This approach enhances the specificity and sensitivity of AAV quantification, ensuring more reliable and reproducible results.
Watch the webinar to learn more about:
- Applying different elution strategies for the baseline separation of empty and full capsids.
- Distinguishing capsid subspecies based on spectroscopic differences in capsid composition.
- Using a light scattering detector for selective analysis of samples with complex matrices and achieving sensitive detection of low-concentration capsid subspecies with fluorescence detector.
- Understanding how post-density gradient ultracentrifugation profiling offers a more affordable and straightforward alternative to analytical ultracentrifugation, eliminating the need for expensive equipment and complex data analysis.
