Ruscic, J;
Perry, C;
Mukhopadhyay, T;
Takeuchi, Y;
Bracewell, DG;
(2019)
Lentiviral Vector Purification Using Nanofiber Ion Exchange Chromatography.
Molecular Therapy: Methods & Clinical Development
, 15
pp. 52-62.
10.1016/j.omtm.2019.08.007.
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Abstract
Lentiviral vectors (LVs) are used in cell and gene therapies due to their ability to transduce both dividing and non-dividing cells while carrying a relatively large genetic payload and providing long-term gene expression via gene integration. Current cultivation methods produce titres of 105 to 107 TU/mL thus it is necessary to concentrate LVs as well as to remove process and product related impurities. In this work we used a packaging cell line WinPac-RD-HV for LV production to simplify upstream processing. A direct capture method based on ion-exchange chromatography and cellulose nanofibers for LV concentration and purification was developed. This novel scalable stationary phase provides a high surface area that is accessible to LV and therefore, has potential for high capacity operation compared to traditional bead-based supports. We were able to concentrate LVs 100-fold while achieving a two-log removal of host cell protein and maintaining up to a 90% yield of functional vector.
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