Three distinct analytical ultracentrifugation methods for virus and viral vector characterizations

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The demands of characterization and quality control of viral vectors and viruses for therapeutic purposes are rapidly increasing. One of the critical quality attributes is a quantitative description of the particle size distribution including analysis of empty, partial, and full particles, as well as aggregates.

Analytical ultracentrifugation (AUC), which was developed for the size distribution analysis of particles in solution by Dr. Theodor Svedberg more than 100 years ago, has been used extensively for the characterization of biopolymers and synthetic polymers. Most recently, modern AUC instruments which utilize direct boundary fitting of sedimentating boundary data have emerged, and the enthusiastic development of analytical software like Sedfit, Ultrascan, DCDT+, SedAnal, and Sednterp has greatly extended the range of applications accessible to AUC.

Sedimentation velocity AUC (SV-AUC) is considered a gold standard method for the size distribution analysis of viral vectors for gene therapy, but several orthogonal AUC methods are appropriate, including band sedimentation AUC (BS-AUC) and density gradient equilibrium AUC (DGE-AUC).

In this webinar, these three AUC methods for the characterization of adeno-associated viral (AAV) vectors will be explained including the principle of each method, examples of data analysis, and the relationship of the results from the three methods.

• The advantages and use cases for SV-, DGE-, and BS-AUC
• Analyzing and interpreting AAV data from each method
• How to utilize each method to obtain a more holistic description of AAV particles

Susumu Uchiyama

Professor, Department of Biotechnology, Department of Engineering, Osaka University

Prof. Susumu Uchiyama is a biophysical chemist with over 25 years of experiences. Since 2017, he is a professor for biotechnology at the Osaka University in Japan. His focus is on the study of solution biophysics of proteins, protein complexes and protein-nucleic acids complexes by using a variety of methods including analytical ultracentrifugation, mass spectrometry (especially native MS and HDX-MS) and calorimetry. Prof. Uchiyama was also involved in remodeling analytical instruments, such as laser optics for Beckman Model-E analytical ultracentrifuge and digital data acquisition system for DASM-4 calorimeter. He has also participated in the development of a number of instrument prototypes. He has published more than 250 peer-reviewed papers and reviews, and edited a book of Analytical Ultracentrifugation published from Springer. He serves on the Editorial Advisory Boards for Journal of Biochemistry and Journal of Pharmaceutical Sciences.

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SPEAKERS

Susumu Uchiyama

Professor, Department of Biotechnology, Department of Engineering, Osaka University

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