CAR-T cell cancer immunotherapy gets personal HarvardResearch NatureComms
T cells in the PBMC sample, were also determined to be important for patient-derived CAR-T cells, consistent with differences observed in our initial blood sample phenotyping. Higher sample numbers would likely improve the accuracy of our prediction model and allow it to describe—given an input T-cell blood sample phenotype—what CAR-T cell products are possible using APC-ms across of a range of stimulation dose.
. MSRs were also evaluated for their ability to coat liposomes and release IL-2, relevant biological proxies to confirm consistent physical structure between batches. MSRs were further confirmed to be endotoxin free via the Limulus Amebocyte Lysate test .Liposomes were prepared as previously described. Lipid films consisting of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine doped with 0.02–0.3 mol% 1,2-di--sn-glycero-3-phosphoethanolamine-- were prepared and dried under nitrogen.
The second-generation CD19-41BBζ CAR construct was composed of the scFv fragment from the FMC63 antibody fused to the human CD8α hinge and transmembrane region and linked to human 4-1BB and human CD3ζ intracellular signaling domains. To enable detection by flow cytometry, a cleavable truncated EGFR was inserted to the N-terminus of the CD3ζ. Lentiviral supernatants were produced using the HEK 293T packaging line as previously described.
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