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. Author manuscript; available in PMC: 2010 Feb 1.
Published in final edited form as: Mol Immunol. 2008 Oct 29;46(5):902–916. doi: 10.1016/j.molimm.2008.09.021

Figure 3. Three different tumor-specific scTCRs stain positive for N- and C-terminal epitope tags, but not Vβ, by flow cytometry.

Figure 3

Yeast transformed with a yeast display plasmid encoding either of three tumor-specific scTCRs (mouse C18 scTCR Vβ-linker-Vα, mWT1-Vα-linker-Vβ, or human CE10 Vβ-linker-Vα) were induced to express the AGA2 fusion. Induced cells were washed and incubated with primary antibody to HA (N-terminal epitope tag), anti-c-myc (C-terminal epitope tag), as well as antibodies to the respective Vβ domains for each single-chain (C18, mouse Vβ8.3; mWT1, mouse Vβ11; CE10, human Vβ21.3) (filled histograms). The Vβ stains were as follows: 4 μg/mL PE-conjugated anti-mouse Vβ8.3 antibody 1B3.3 for the C18 scTCR, 4 μg/mL PE-conjugated anti-mouse Vβ11 antibody RR3-15 for the mWT1 scTCR, and 5 μg/mL anti-Vβ21.3, followed by biotin goat anti-mouse and SA:PE for the CE10 scTCR. Samples of induced cells were incubated with secondary antibody alone (grey outline).