Label-free purification of viable human T-lymphocyte cells from a mixture of viable and non-viable cells after transfection by electroporation

Recent developments in chimeric antigen receptor (CAR) T-cell therapy for cancer have shown promising results. CAR T-cells are engineered mainly by AC or DC electroporation to express CAR molecules on their surfaces. Electroporation has detrimental effects on the majority of T-cells and causes cell death via apoptosis or necrosis. These non-viable cells have potentially harmful effects on other healthy T-cells. Because there are no unique biomarkers that pinpoint the apoptosis and necrosis of T-cells, currently available cell separation methods are incapable of purifying healthy T-cells after electroporation. To address this critical issue, we have used dielectrophoretic cell isolation in a simple microfluidics chip. Our results indicate that it is possible to purify cell samples after electroporation and achieve ~100% purity and??>90% target cell recovery. Therefore, this work presents a viable solution to a critical need in CAR T-cell manufacturing.