Second-generation CD33-CAR incorporating the My96 scFv sequence was constructed as described earlier [22 (link)]. Briefly, the My96 scFv was combined in frame with CD8 hinge and transmembrane domain, 4-1BB/CD137 co-stimulatory domain, and CD3ζ activation domain. A leader peptide derived from GM-CSFRα was included to facilitate CAR cell surface expression. Third-generation self-inactivating baboon envelope-pseudotyped lentiviral vectors (BaEV-LVs) were produced by transient transfection into HEK293T cells using MACSfectin (Miltenyi Biotec) or polyethylenimine (PEI) [23 (link)].
Albinger N., Pfeifer R., Nitsche M., Mertlitz S., Campe J., Stein K., Kreyenberg H., Schubert R., Quadflieg M., Schneider D., Kühn M.W., Penack O., Zhang C., Möker N, & Ullrich E. (2022). Primary CD33-targeting CAR-NK cells for the treatment of acute myeloid leukemia. Blood Cancer Journal, 12(4), 61.
Other organizations :
Goethe University Frankfurt, Charité - Universitätsmedizin Berlin, Freie Universität Berlin, Humboldt-Universität zu Berlin, Frankfurt Cancer Institute, Lentigen Technology (United States), University Medical Center of the Johannes Gutenberg University Mainz, Johannes Gutenberg University Mainz, University Hospital Frankfurt
Second-generation CD33-CAR incorporating the My96 scFv sequence
dependent variables
CAR cell surface expression
control variables
Leader peptide derived from GM-CSFRα included to facilitate CAR cell surface expression
controls
Positive control: Not explicitly mentioned
Negative control: Not explicitly mentioned
Annotations
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