Scalable Non-Viral CAR-NK Cell Manufacturing Using the TcBuster™ Transposon System

Advancements in gene delivery technologies have significantly enhanced the ability to engineer immune cells for cancer immunotherapy. TcBuster, a non-viral transposon-based gene delivery system, enables stable integration of therapeutic genes into natural killer (NK) cells and T cells, to support the development of CAR-NK and CAR-T cell therapies. 

This application note highlights a fully scalable CAR-NK cell manufacturing workflow using the TcBuster transposon system, ExCellerate™ Human NK Cell Expansion Media, and the Neon NxT and CTS™ Xenon Electroporation Systems. Gene delivery to NK cells was optimized at a small scale using the Neon NxT Electroporation System and the process was translated to clinical-scale production by applying consistent parameters on the Xenon Electroporation System. This workflow delivered high transposition efficiencies and cell viability across diverse donors, establishing an efficient, reproducible process for CAR-NK cell production.

Key Takeaways

• Combined use of the TcBuster-M Transposon system, ExCellerate Human NK Cell Expansion Media, and the Neon NxT and Xenon Electroporation Systems enables a robust, scalable workflow for the generation of CAR-NK cells.
• Consistent electroporation parameters allow a smooth transition from research-scale optimization on the Neon NxT System to clinical-scale manufacturing on the Xenon System.
• The use of TcBuster with either electroporation system delivers high transposition efficiencies, coupled with high cell viability and expansion rates.
• The workflow outlined allows an average of 1.3 x 10¹¹ CD19-CAR-NK cells to be manufactured with the research-scale Neon NxT System, while an average of 7.2 x 10¹¹ CD19-CAR-NK cells can be manufactured using the clinical-scale Xenon System.