AAV capsid analysis and quantification with chromatography and ultracentrifugation-based techniques

Thursday 08:00 PST / 11:00 EST / 16:00 GMT / 17:00 CET

Sponsor

AAV capsid analysis and quantification with chromatography and ultracentrifugation-based techniques

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.

Andreja Livk

Head of Process Analytics at Sartorius BIA Separations

Andreja is a Head of Process Analytics Development department at Sartorius BIA Separations. She leads process analytics development team that focuses on the further advancement of a chromatographic solutions, involving the development of innovative chromatographic analytical methods for the analysis of biological molecules, including mRNA, pDNA and viruses. Over the course of career with biotech companies, she gained a wide range experience in protein chemistry, including characterisation of biosimilars and biologics. She holds a Master Science in Biochemistry from the University of Ljubljana, Slovenia.

Sebastijan Peljhan

Head of Basic Research at Sartorius BIA Separations

Sebastijan is a Head of Basic Research department at Sartorius BIA Separations. His group focuses on research and development of new monolith based applications, including immobilized enzymatic reactors and affinity chromatography columns. The team also developed post-DGUC profiling approach for characterization of large biomolecules.

Sebastijan holds master’s degree in physical chemistry from the University of Ljubljana, Slovenia and he completed his PhD in theoretical chemistry at Jožef Stefan Institute in Ljubljana, Slovenia. He has a wide range of experiences in chromatographic purification and in-process chromatographic characterization of blood plasma proteins, viruses and nucleic acids.

in 15

Days