Enrichment of full rAAV capsids in a scalable, reproducible viral vector manufacturing platform

Recombinant adeno-associated viruses (rAAV) are the gene transfer vector of choice for many in vivo therapies. These vectors synthetic viral particles which can deliver a therapeutic to patient or patients’ cells correct genetic abnormality. be produced in single-use bioreactors and purified using scalable technologies. We evaluated use scalable, filtration chromatography technologies downstream purification an rAAV5 vector. In this testing, was Pall iCELLis® Nano bioreactor by polyethylenimine (PEI) mediated triple-plasmid transfection. The harvest material clarified direct flow with combination Seitz-P grade depth 0.2 mm sterilizing filters. product concentrated 100 kDa OmegaTM Membrane flat-sheet tangential flow-filtration (TFF) before primary affinity chromatography. Affinity polished Mustang® Q membrane enrich full capsids. then diafiltered final formulation Omega TFF membrane. sterile filtered Pall’s Supor® EKV validated sterilizing-grade This manufacturing process optimized yield, low contaminant profile capsid enrichment. established feasibility near complete end-to-end almost all materials available from Corporation. resulted theoretical whole yield ~25% (host cell protein [HCP] [DNA]) ~5-fold enrichment capsids total described here shows potential platformable rAAV products.