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. Author manuscript; available in PMC: 2019 Sep 4.
Published in final edited form as: Nat Biomed Eng. 2019 Mar 4;3(4):292–305. doi: 10.1038/s41551-019-0360-0

Figure 1. αPD-1-ABD-PE consisting of αPD-1, ABD, and PE specifically binds with and enters PD-1+ lymphocytes.

Figure 1.

(a) The sequential configurations of the functional domains, αPD-1, ABD, and PE, in αPD-1, ABD-PE, αPD-1-PE, and αPD-1-ABD-PE. The linker, (GGGGS)3, is shown as an orange box. (b) The amino acid sequences of αPD-1 VH and VL with their framework regions (FRs) and complementarity-determining regions (CDRs) highlighted with red text. Two mutations were introduced in the VH and VL, respectively. The two mutations are underlined, VH: R45C; VL: G104C. (c) The mean fluorescence intensity (MFI) of PD-1+ and PD-1− primary T cells after the cells were incubated with Alexa Fluor 647-labeled αPD-1-ABD-PE or Alexa Fluor 647-labeled ABD-PE at 4°C for 30 minutes. The cells were collected from C57BL/6 mice. The MFI means and their standard deviations (SDs) are indicated (N=6 biologically independent samples; unpaired two-sided t-test). The MFI was obtained by flow cytometry. (d) The MFI of PD-1+ and PD-1− primary B cells after the cells were incubated with the labeled αPD-1-ABD-PE or the labeled ABD-PE at 4°C for 30 minutes. The cells were collected from C57BL/6 mice. The MFI means and their SDs are indicated (N=6 biologically independent samples, unpaired two-sided t-test). (e) The MFI of PD-1+ and PD-1− primary T cells after the cells were incubated with the labeled αPD-1-ABD-PE or the labeled ABD-PE at 37°C for 30 minutes. The cells were collected from C57BL/6 mice. The MFI means and their SDs are indicated (N=6 biologically independent samples; unpaired two-sided t-test). (f) The MFI of PD-1+ and PD-1− primary B cells after the cells were incubated with the labeled αPD-1-ABD-PE or the labeled ABD-PE at 37°C for 30 minutes. The cells were collected from C57BL/6 mice. The MFIs and their SDs are indicated (N=6 biologically independent samples; unpaired two-sided t-test). (g) The MFI of EL4 cells after the cells were incubated with the labeled αPD-1-ABD-PE under the conditions noted in the figure. The MFIs and their SDs are indicated (N=6 biologically independent samples; unpaired two-sided t-test). Studies described in the panels c-g were repeated twice with similar results. Data of one repeat is shown here.