Abstract: Provided herein are methods for producing genetically engineered immune cells, such as T cells. In some aspects, the immune cells are genetically engineered by targeted integration of a transgene into a target site of a gene in the immune cells. In some embodiments, the genetically engineered immune cells are produced from a whole blood sample. In some aspects, the immune cells are genetically engineered following on-column stimulation of the immune cells. In some aspects, the immune cells are genetically engineered by non-viral gene delivery methods. Also provided herein are related cells, compositions, and uses.

Abstract: Provided herein are methods for selecting and stimulating a plurality of cells in a sample of cells using column chromatography, and collecting the cells without using additional steps or reagents to facilitate detachment of the cells from the column. In some aspects, the methods provided herein reduce the time needed to generate a population of selected and stimulated cells useful for genetic engineering, and ultimately, cell therapy, compared to existing methods. Also provided are articles of manufacture and apparatus thereof.