Fig. 2
ROR1/CD3 TCE shows high binding affinity to ROR1 and CD3. (A) The schematic diagram of ROR1/CD3 antibody. The green part is the Fab targeting ROR1, and the right yellow part is the single-chain variable fragment (scFv) targeting CD3. ROR1/CD3 TCE molecule consists of three chains: anti-ROR1 antibody heavy chain (chain 1), anti-ROR1 antibody light chain (chain2) and an anti-CD3 antibody ScFv-Fc fused chain (Chain 3). (B, C) Cell-based binding of ROR1/CD3 TCE molecule to ROR1-expressing MDA-MB-231 cell and CD3-expressing human T cells. Cells were incubated with a serially diluted ROR1/CD3 antibody followed by staining with PE-conjugated anti-human IgG. The MFI was determined by flow cytometry. The experiment was performed with three technological replicates and validated by other two biological replicates. (D) The binding affinity of ROR1/CD3 to ROR1 and CD3 was evaluated by the surface plasma resonance. The experimemnt was performed with three technological replicates. KD, binding dissociation equilibrium constants; Ka, kinetic association rate; Kd, kinetic dissociation rate