Study design and CAR-37 T-cell manufacturing. (A) Design of the lentiviral vector construct used to produce CAR-37 T cells. (B) Study design and (C) CAR-37 manufacturing process. (D) End-product percentage transduction of patient T cells as measured by the percentage EGFR⁺CD3⁺CD45⁺ cells in the live cell population (by 4′,6-diamidino-2-phenylindole staining) via flow cytometry. (E) Copies of CAR-37 vector per microgram of DNA in the final product as measured by PCR. (F) The total number of CAR-37 T cells produced per patient and available for dosing (left y-axis, solid bars) and the fold expansion of the patient’s T cells during manufacturing (x-axis, striped bars), along with the dose of CAR-37 T cells they received (black dots), are shown. (G) Percent of the final product that is CD4⁺ (colored bar, bottom) vs CD8⁺ (white bar, top). Cy, cyclophosphamide; DP, drug product; flu, fludarabine; PBMC, peripheral blood mononuclear cell; scFv, single-chain variable fragment.