Chimeric antigen receptors (CARS) targeting hematologic malignancies, compositions and methods of use thereof
A chimeric antigen receptor and antibody technology, applied in the direction of antibody mimics/scaffolds, antibody medical components, DNA/RNA fragments, etc.
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[0387] Production of the third generation CD5CAR
[0388] Constructs of CD5CAR and anchored CD5 scFv antibodies were designed to test the function and mechanism of CD5CART cells to target and lyse CD5 expressing cells and the ability of CD5CART cells to downregulate CD5 expression within their own CD5CART cell population ( Figure 17A ). To confirm the CD5CAR construct, the resulting CD5CAR lentivirus was transduced into HEK293 cells. After being treated with CD5CAR or GFP-lentivirus for 48 hours, the expression of CD5CAR in HEK293 cells was verified by Western blot analysis using CD3ζ antibody, which recognizes the C-terminus of CD5CAR protein ( Figure 17B ). The resulting band was the expected size of CD5CAR protein in CD5CAR-transduced HEK293 cells, but GFP-transduced HEK293 cells did not show any specific bands by western blot analysis. To evaluate the function of CD5CAR protein for subsequent experiments, CD5CAR lentivirus was transduced into activate...
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