Figure 3.
αFAP-E5B9L TMs expression and binding assessment to UniCAR T-cells using flow cytometry. TMs were produced recombinantly by stable producer cell lines and purified from cell culture supernatants using His-tag affinity chromatography. (A) Purified TMs were analyzed using SDS-PAGE under reducing conditions followed by Coomassie staining. (B) Alternatively, the separated proteins were transferred to nitrocellulose membrane and detected by immunoblotting using anti-penta-His mAb. (C) The binding of different concentrations of αFAP TMs to UniCAR T-cells was detected using anti-His antibody and flow cytometry. Statistical analysis was performed using GraphPad Prism 9.0 and statistical significance was determined using two-tailed paired T-test (* p ≤ 0.012). M: molecular weight. KD: equilibrium dissociation constant.