Scaling Up Non-Viral CAR-T Cell Manufacturing Using the TcBuster™ Transposon System

Advancing CAR-T cell therapies requires gene editing technologies that are both efficient and scalable. TcBuster, a non-viral transposon-based gene editing system, offers a robust solution for stable therapeutic gene integration into primary human T cells using standard electroporation methods.

In this study, we demonstrate the ability to efficiently generate CD19-CAR-T cells from both pre-activated and resting T cells using the TcBuster transposon system with the Neon NxT and CTS™ Xenon Electroporation Systems. The Neon NxT Electroporation System was used for small-scale optimization of gene delivery, and the process was translated to a clinical-scale workflow by applying consistent parameters on the Xenon Electroporation System. The use of TcBuster with the Xenon System produced viable, genetically edited T cell quantities well above typical CAR-T cell dosing requirements, underscoring the potential of this integrated, closed system compatible workflow for scalable, GMP-compliant CAR-T cell manufacturing.

Key Takeaways

• Combined use of the TcBuster-M Transposon system and the Neon NxT and Xenon Electroporation Systems enables a robust, scalable workflow for the generation of CAR-T cells.
• Consistent electroporation parameters allow a smooth transition from research-scale optimization on the Neon NxT Electroporation System to clinical-scale manufacturing on the Xenon System.
• The use of TcBuster with either electroporation system delivers robust transposition efficiencies, exceeding 30% in activated T cells and 15% in resting T cells, accompanied by high cell viability and expansion rates exceeding 25-fold.
• Employing TcBuster with the Xenon Electroporation System generates 1.2 billion viable, genetically edited CD19-CAR-T cells from pre-activated T cells and 800 million from resting T cells across three donors, exceeding dosing requirements for current FDA-approved CAR-T cell therapies.