Non-Viral Delivery Of Complex Cargos For Large-Scale T Cell Manufacturing

By Frank Morrissey-Wettney, Hannah Goldfarb Wittkopf, Nadessa Poeppel, Meike Zander, Nicole Kukli, Katharina Stolle, Gina Andretta-Beu, Stefanie Müthel, Valeria Annibaldi, and Andrea Toell

Non-viral cell engineering techniques have gained prominence as promising substitutes for viral transduction. Among vector-free approaches, electroporation stands out as the gold standard and is increasingly utilized in clinical trials. We engineered an enhanced cartridge for the 4D-Nucleofector® LV Unit PRO to facilitate the electroporation of complex molecules into large T cell populations. Initially, we optimized cell handling parameters using 100 µL reactions before scaling up to transfect up to 1×10⁹ cells in 10-20 mL with the new large-scale cartridge.