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Proteins, Expression, Isolation and Analysis> | … reported (Zheng et al., 2012). Briefly, 1 × 106 CAR-T cells were suspended in 0.2 mL ice-cold wash buffer (PBS containing 2% BSA) with 0.02 µg protein L (GenScript, Cat. No. M00097) for 30 min. After washing three times, CAR … | Get A Quote |
Chimeric antigen receptor T (CAR-T) cell therapy is not satisfying in solid tumors. PD-1-mediated suppression greatly hinders CAR-T cells in the microenvironment. It has been shown that PD-1 blockade improves the effectiveness of CAR-T cells. Herein, we designed CAR-T cells than could secret α-PD-1 scFv by themselves. To obtain optimal secretions of scFv, we screened several signal peptides. And the segment from human increased the extracellular production of PD-1-neutralizing proteins. The secreted neutralizing scFv efficiently blocked PD-1 and enhanced T cell activation when PD-L1 was present. Further analysis showed that CAR-T cells themselves could secret α-PD-1 scFv with bioactivity. In contrast to the p... More