Abstract
AbstractAutologous cellular therapies have been highly successful in treating hematological cancers and have the potential to be used for a variety of indications. Manufacturing these therapies rapidly and at low cost remains a major challenge. A key bottleneck in cellular therapy manufacturing is genetic modification of target cells, which is often done using viral vectors. Because vectors are expensive to develop and produce, non‐viral gene transfer using electroporation is emerging as a preferred transfection method for next‐generation therapies. However, most commercial electroporation systems are built for research use rather than large‐scale clinical manufacturing. The microfluidic, continuous‐flow electroporation device presented here offers several advantages including large‐scale and high throughput processing, high performance, and the potential for automation. It transfects primary human T cells with Cas9‐guide ribonucleic acid (RNA) ribonucleoprotein complexes (RNP) and messenger RNA (mRNA) with up to 99–100% efficiency and minimal impact on viability. In addition, this device transfects 3.5 kbp plasmid deoxyribonucleic acid with up to 86% efficiency after preliminary optimization studies. A single microchannel can deliver a total cellular processing throughput of up to 9.6 billion per hour. The combination of high throughput and high performance enables the scale of processing required for future “off‐the‐shelf” allogeneic cellular therapies.
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