Enhanced AAV Purification Using Choline Chloride

This technical report presents a novel method for the purification of adeno-associated virus (AAV) using an optimized anion exchange chromatography (AEX) process. The method utilizes choline chloride as the eluting salt, which significantly improves the separation of full AAV capsids from empty capsids. The report outlines the limitations of traditional preparative ultracentrifugation, including its time-intensive nature and scalability issues. The AEX method described is non-toxic, scalable, and adaptable to various AAV serotypes, achieving a full AAV content of 93%, comparable to ultracentrifugation. Experimental conditions for the AEX process are detailed, including the use of a SkillPak TOYOPEARL GigaCap Q-650S column and specific mobile phase compositions. The results demonstrate the effectiveness of the choline chloride-based method in enhancing full AAV capsid enrichment while reducing operational costs and time. This report serves as a guide for researchers and professionals in the field of gene therapy and biopharmaceuticals.