Downstream Processing

Disruptive bench-scale purification of lentivirus using affinity-liquid phase separation technology

Cell & Gene Therapy Insights 2023; 9(9), 1255–1269

DOI: 10.18609/cgti.2023.163

Published: 1 November
Nicole L Votaw, Melissa Callander, Torie Broer, Alyssa Wheeler, Michael Dzuricky, Kelli Luginbuhl

Lentiviral vectors are widely used in cell and gene therapy, but their high production costs hamper early-stage research efforts. Furthermore, many concentration and purification methods available to early-stage researchers today are difficult to scale or lack specificity for lentiviral purification, hindering preclinical research and translation to clinical and commercial scales. We have developed a research-scale lentiviral reagent with the potential to transform lentiviral purification workflows. The reagent is quick and easy to use, yields high titer lentiviral vectors, and, because of its specificity, is more effective at contaminant removal than other LV purification products designed for research use. By specifically attaching to the viral envelope in solution and then forming liquid droplets around the lentivirus, IsoTag™ LV protects viruses from aggregation and degradation while dramatically increasing the effective size and density of the LV, thus facilitating easy separation from other smaller and less dense contaminants (i.e., host cell proteins). The actual process of capture is very simple and can be implemented by any user, at any skill level, with low-cost lab equipment. The design and development of a small-scale centrifugation process and novel reagent detailed herein lays the foundation for additional development of the IsoTag™ LV for larger scale processing. IsoTag™ LV is poised to democratize LV production and purification in the research world and accelerate development of new cell and gene therapies.