Adeno-associated virus (AAV) is a go-to vector for gene therapy treatments. In each AAV downstream process, one of the key steps (and often a bottleneck) is enrichment of full capsids. Along with scalability and capsid separation challenges, one must account for sample heterogeneity and assess which analytics are adequate to distinguish between AAV subpopulations and deliver only the potent AAV product.
Chromatography on monolith columns offers efficient and scalable downstream processes. Paramount considerations for scaling up will be discussed and a case study will be presented using a Design of Experiments approach for buffer selection and robust preparation for improved separation of empty and full capsids. In addition, residual host cell proteins, host cell DNA, plasmid DNA, and endotoxin removal efficiency will be discussed.