Isolation of Peripheral Blood Mononuclear Cells for Tisagenlecleucel CAR T-Cell Therapy

Peripheral blood or bone marrow was collected into EDTA tubes from patients receiving tisagenlecleucel CAR T cell therapy as part of their standard of care for routine clinical testing. Isolation of the mononuclear cells (MNCs) was performed using a density gradient with centrifugation [15 (link)] (STEP 4), and DNA was extracted from the MNCs using the QIAamp DNA Blood Mini Kit as described previously [18 (link),25 (link),26 (link)] (Qiagen, Hilden, Germany). Extracted DNA was quantified using the Nanodrop 1000 (Labtech Ltd., East Sussex, UK) and diluted to 10 ng/µL (STEP 5). When CD3 T cells were isolated, Miltenyi Biotec microbead kits were used.