Fig. 1
Specific binding capacities of CD3-HAC to PD-L1-positive cells and CD3-positive cells. a Schematic representation of adenoviral expression vector for CD3-HAC. hTERT, promoter of human telomerase reverse transcriptase; SP, signal peptide, a murine kappa light-chain leader peptide; His6, hexa-histidine tag; G4S, Gly-Gly-Gly-Gly-Ser residues. b, c PD-L1 expression on breast cancer cells (MDA-MB-231 and MCF-7) detected by flow cytometry and immunofluorescence. (a) Negative control. (b) PE-labeled anti-PD-L1 antibody. Blue (nuclei); red (anti-PD-L1). Scale bar, 10 μm. d Competitive binding activity of purified CD3-HAC with anti-PD-L1 antibody on MDA-MB-231 cells. (a) Negative control. (b) CD3-HAC+anti-PD-L1 antibody. (c) Anti-PD-L1 antibody alone. e Binding specificity of CD3-HAC to Jurkat cells. (a) Negative control. (b) CD3-HAC. f, g Flow cytometry and immunofluorescence analyses performed on breast tumor cell lines infected by AdCD3-HAC to detect the binding specificities of CD3-HAC with anti-His antibody. Blue (nuclei). Green (GFP) indicates the cells infected by adenovirus. Red (anti-His for CD3-HAC). Scale bar, 35 μ m