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. Author manuscript; available in PMC: 2021 Apr 8.
Published in final edited form as: J Am Chem Soc. 2020 Mar 30;142(14):6554–6568. doi: 10.1021/jacs.9b11622

Figure 4.

Figure 4.

Simultaneous arming of SpyCatcher T cells with two targeting ligands generates a single cell product capable of dual antigen targeting. (A) Schematic representation of single vs dual targeting ligand loading. (B) SpyCatcher T cells were armed with either 1000 nM myc-9.26-ST (αHer2; red), 1000 nM Flag-E01-ST (αEGFR; blue), or both simultaneously at 1000 nM each (green). Receptor loading was detected with a combination of fluorescently conjugated anti-myc and anti-flag antibodies and assessed via flow cytometry. (C) Single- or dual-armed SpyCatcher T cells were cocultured with Ramos cells expressing either Her2 or EGFR and luciferase. Residual luciferase expression was calculated after 20 h. All cocultures were carried out using a 7:1 E/T ratio. Data points are averages of three replicates. A representative T-cell donor is shown.