Want to improve lentiviral vector yields?

Robert Lou, PhD

Robert Lou, PhD

Associate Director

Exploring Innovative Methods for Enhancing Lentiviral Vector Yield and Target Cell Infectivity in CAR-T Cell Therapy Manufacturing


In recent years, labs and companies have been seeking ways to reduce the cost of manufacturing CAR-T products to increase the accessibility of the CAR-T treatment. Advances in cell processing and manufacturing techniques have improved in recent years. Along with technology improvements, the Theragent viral vector lab is continuously exploring novel methods for enhancing the efficiency and quality of the procedures in CAR-T manufacturing and herein we will demonstrate successful achievement of these goals.

First, our lab has developed several new viral production cell lines and a modified traditional transduction method for the viral production and cell modification. By using the newly developed production cell line, the yield of the viral vector can be significantly increased at least 5-fold in unprocessed viral titer compared to the traditional adherent method and additionally, at least 25% of the production yield compared to the current suspension culture lines available on the market. In addition, using the modified cell line we found that the capacity of the gene insertion size in the testing viral transfer plasmid can be significantly increased to 6~8 KB CDS length while maintaining its related production titer. This provides more flexibility in the vector design. Our lab has also developed a method that enables improvement in the infectability of cells using a significantly lower viral MOI during the CAR-T development process. With the designed drug treatment, treated cells can be sensitized for infection with 3-fold lower MOI and with relatively higher expression of the genes of interest. This improvement allows the manufacturer to produce CAR-T cells in a more cost-effective manner. Meanwhile, through collaborative efforts, we are currently developing new platforms for generating safe tools for CAR-T development by using CRISPR technology and the self-inactivated retro-viral vector. This collaboration will focus on the improvement in the safety and efficiency of the procedures. Combining our several improvements, the result shows our method can be applied into current CAR-T production procedure and with a significant positive impact to the operational timing and cost. In addition, our results also suggest that the on-current procedure may offer potential for further improvement in the future.

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