Development of large-scale downstream processing for lentiviral vectors

Abstract The interest in lentiviral vectors has increased prominently for gene therapy applications, but few have reached the later stages of clinical trials. The main challenge has remained in scaling up the manufacturing process for the fragile vector to obtain high titers for in vivo usage. We have previously scaled up the LV production to iCELLis 500, being able to produce up to 180 liters of harvest material in one run with perfusion. The following challenge considers the purification and concentration of the product to meet titer and purity requirements for clinical use. We have developed a downstream process beginning with clarification, buffer exchange and concentration by tangential flow filtration. This is followed by a purification step using single membrane-based anion exchange chromatography and final formulation with tangential flow filtration. DiffeThe concentration and purification of the LVs was optimized and scaled up to process a 180 L harvest, with the main optimization concentrating on anion exchange chromatography. The process yielded a high-titer LV product, with majority of the impurities removed.