Deciphering Empty-Full Characterization: A Comparative Analysis of Available Analytical Methods

Empty full ratio of AAV particles in a gene therapy drug is an important critical quality attribute. Due to the complexity of biological production process specifically, viral vector production mechanisms, AAV particles are quite heterogeneous. Purification process can’t entirely separate empty, partial and full particles from the process and hence a significant amount of empty and partial particles gets retained in the final product, potentially compromising quality of the product and raising safety concerns. The complexity of the challenge is understandable as the as gene therapy drugs contain multimeric protein assembly enclosing a genetic material. Where the actual process of DNA packaging remains loosely understood. There are various analytical technologies available for the characterization of these particles but almost every technology has some limitations, either in terms of the sample requirement, quality of information, precision, affordability, complexity of the assay or for their suitability in the GMP environment. AUC maintains its central role in this process and remains as a gold standard but at the same time has limited applicability across various phases of drug development due to its limited throughput and high sample requirements. There are various alternative technologies coming into play, including DLS, CDMS, Mass Spectrometry, Mass Photometry, ddPCR/Capsid ELISA, AEX chromatography, Cryo-TEM and UV-Vis measurements. In this webinar we will discuss opportunities and challenges in various analytical technologies in characterizing this critical quality attribute of AAV vector for gene therapy.

Presented by Binod Pandey, Senior Scientist at Novartis

Dr. Binod Pandey received his PhD from the University of Missouri-St Louis,USA and completed postdoctoral work at Pennsylvania State University. His expertise spans from analytical chemistry, separation science, biochemistry and surface chemistry. He has held various roles in pharmaceutical industries in analytical departments for supporting process and product development and characterization, in every phase of the drug development. His industry expertise involves analytical characterization of various biologics including proteins, fusion proteins, mAb, antibody drug conjugates (ADCs) and viral vector characterization (AAV and LVV). As a Senior scientist at Novartis technical research and development division at cell and gene therapy unit, he has led various projects in viral vector characterizations, including capsid and genome characterization, impurity analysis and empty full characterization using AUC and orthogonal methods. He has published many papers and presented in various national and international conferences. He is an expert in analytical characterization of AAV including empty, partial and full characterization, and holds the expert knowledge on current opportunities and challenges in the field.