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. 2023 May 25;14:1182409. doi: 10.3389/fimmu.2023.1182409

Figure 3.

Figure 3

Comparison of the detection and expression of several scFvs CAR by flow cytometry. (A) The expression of the 4 CAR scFvs on the same cells was evaluated on days 6, 10, and 14 of cell culture using three methods: CEA-His, protein L and GFP. The values are presented as the mean ± SD of four independent experiments derived from 4 different donors (n=4). (B) When CAR expression was detected using CEA-His, the results for the 4 types of CAR-T cells were compared. hMN-14, BW431/26 and C2-45 CAR-T cells were compared to M5A cells. The values are presented as the mean ± SD of four independent experiments (n=4). (C) Representative FCM results for protein L and CEA-His in the 4 types of CAR-T cells. (D) The MFI of protein L and CEA-His was also measured and compared on day 10 of cell culture. The data are presented as the mean ± SD of five independent experiments (protein L) (n=5) and four independent experiments (CEA-His) (n=4). (E) HEK293T cells were transduced with each CAR viral vector, and CAR expression was measured by assessment of protein L and CEA-His. Representative FCM results from three independent experiments are shown (n=3). Statistical analysis was performed by a paired t-test. * = p < 0.05; ** = p < 0.01; and ns, not significant.