Texmacs t cell isolation3/1/2023 1–4 At present, ‘conventional’ CAR T cell manufacturing takes place in labor-intensive production campaigns that entail a complex sequence of manual and ‘open’ process steps, with little to no automation. Immunotherapy with chimeric antigen receptor (CAR) T cells has demonstrated therapeutic potential for treating B cell leukemia and lymphoma and multiple myeloma, resulting in a steadily increasing demand for CAR T cell products by caregivers and patients. Notably, the operator hands-on-time was substantially reduced compared with conventional non-automated CAR T cell manufacturing campaigns. The CD19 CAR T cell product conferred potent anti-lymphoma reactivity in pre-clinical models. The transposon copy number was 7.0, 9.4 and 6.8 in runs #1–3, respectively, and gene analyses showed a balanced expression of activation/exhaustion markers. The resulting cell product contained highly pure T cells (97.3☑.6%) with balanced CD4/CD8 ratio and a high frequency of T cells with central memory phenotype (87.5%☑0.4%). ![]() In three large scale runs, 2E8 T cells were enriched from leukapheresis product, activated, gene-engineered and expanded to yield up to 3.5E9 total T cells/1.4E9 CAR-modified T cells within 12 days (CAR-modified T cells: 28.8%☑2.3%). Results We defined a bi-pulse electroporation shock with bi-directional and unidirectional electric field, respectively, that permitted efficient MC insertion and maintained a high frequency of viable T cells.
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