1 Department of Electrical and Computer Engineering, Concordia University, Montréal, Québec H4B 1R6, Canada.
2 Centre for Applied Synthetic Biology, Concordia University, Montréal, Québec H4B 1R6, Canada.
3 Department of Chemical Engineering, Concordia University, Montréal, Quebec H4B 1R6, Canada.
4 Department of Biology, Concordia University, Montréal, Québec H4B 1R6, Canada.
5 Institut de Recherche en Immunologie et en Cancerologie, Université de Montréal, Montréal, Quebec H3T 1J4, Canada.
6 Drop Genie, Inc., Boston, Massachusetts 02111, United States.
7 Département de Pathologie et Biologie Cellulaire, Université de Montréal, Montréal, Quebec H3T 1J4, Canada.

Genetically engineering human immune cells has been shown to be an effective approach for developing novel cellular therapies to treat a wide range of diseases. To expand the scope of these cellular therapies while solving persistent challenges, extensive research and development is still required. Here we use a digital microfluidic enabled electroporation system (referred to as triDrop) specifically designed to mitigate harm during electroporation procedures and compare against two state-of-the-art commercially available systems for the engineering of primary human T cells. We describe the ability to use triDrop for highly efficient transfection with minimal reagent consumption while preserving a healthy transcriptomic profile. Finally, we show for the first time the ability to use a digital microfluidic platform for the miniaturized production of Chimeric Antigen Receptor (CAR) T cell therapies demonstrating how this novel system can lead to a 2-fold improvement in immunotherapeutic functionality compared to gold standard methods while providing up to a 20-fold reduction in cost. These results highlight the potential power of this system for automated, rapid, and affordable next-generation cell therapy R& D.