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The vector of anti-CD123 chimeric antigen receptor (CAR) is constructed for the engineering of T cells to target Human CD123. The T cells are genetically modified through transduction with a lentiviral vector expressing scFv of anti-CD123 antibody linked to CD28 and CD3ζ signaling domains. And the vector product was designed for the treatment of Acute myeloid leukemia (AML).
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CAR Construction :
Fig.1 Cell surface-binding of 12F1 scFv. Binding of 1 mg/ml of 12F1 to CHO-CD123 was analyzed by flow cytometry with PE-conjugated secondary antibodies. Kuo, S. R., Wong, L., & Liu, J. S. (2012). Engineering a CD123xCD3 bispecific scFv immunofusion for the treatment of leukemia and elimination of leukemia stem cells. Protein Engineering, Design & Selection, 25(10), 561-570. |
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CAR Construction :
Fig.2 Cell-based ELISA binding of 12F1 scFv. 96-well plate coated with CHO-CD123 cells/well was fixed and probed by serial titrations of 12F1 (square) and detected with HRP-conjugated goat-anti-mouse (12F1) antibodies. The average of three sets of reactions was plotted by GraphicPad Prism. Kuo, S. R., Wong, L., & Liu, J. S. (2012). Engineering a CD123xCD3 bispecific scFv immunofusion for the treatment of leukemia and elimination of leukemia stem cells. Protein Engineering, Design & Selection, 25(10), 561-570. |
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