Gene and cell therapies have the potential to dramatically expand the treatment possibilities for patients with conditions that have no other cures. Viral vectors, and particularly lentiviral vectors (LVs), are among the fastest growing types of gene-delivery vehicles. However, scalable production of LVs with high purity and high yield is a major challenge due to the fragility of LVs and a lack of efficient purification tools.Robust affinity chromatography products are desired to simplify the purification process and increase recovery. Here, we present a new affinity chromatographic membrane technology for the rapid purification of LVs with high binding capacity at short residence time and high recovery using mild elution conditions. <br/>The LVs were grown from 293FT cells, and the culture supernatant with a virus titer of approximately 10^6 virus particles per mL was loaded directly onto Purilogics scaled-down syringe filter-like affinity membrane column prototypes. The affinity interaction is based on reversible binding of the LV envelope protein. LVs were eluted under mild conditions to enable high recovery of active virus. Activity was assessed by transduction efficiency of green fluorescent protein (GFP) into HEK293 cells. LV purity was determined by SDS-PAGE and ELISA assays. Additionally, we will show a side-by-side performance comparison to ion-exchange membranes in terms of binding capacity, yield, and purity.<br/>

Affinity membrane for rapid, high-recovery concentration and purification of lentivirus vectors

ACS Fall 2020 Virtual Meeting

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