A Chimeric Approach To Purifying Lentiviral Vectors For CAR-T

Lentiviral vectors (LVV) are the most common delivery method for transducing T cells for CAR-T therapies, and their production and purification are major cost-drivers in manufacturing; an industry “gold standard” of 10-20% recovery results in oversized and expensive production batches. Effective and consistent purification strategies for LVV are a serious challenge due to 1) the labile nature of the virus, 2) physically segregating LVV from the cells from which they bud, 3) removing host cell DNA and protein, and 4) 0.2 μm sterile filtration for 0.08-0.12 μm particles at high concentrations. Although unit operations from mAB and vaccine bioprocessing are readily available, they have yet to be successfully commercialized for LVV (e.g. affinity chromatography) or are detrimental to infectivity (e.g. anion exchange). Thus, a host of wildly divergent, open, and non-scalable schemes are being developed across industry and academia, resulting in poor recoveries, inconsistency of product, and risk of contamination.

Although much work needs to be done to fully optimize each unit operation, we present here our results to date in designing a fully scalable, single-use consumables, closed, lentiviral vector purification process.