Fig. 6

The anticancer effect of anti-CD47/CD20 bispecific antibody against lymphoma in vivo. a The structure of anti-CD47/CD20 bispecific antibody. b The reductive (left) and nonreductive form (right) of BsAb were displayed through SDS-PAGE. c The purity of BsAb was determined through HPLC analysis. The above experiments were performed in triplicate and one representative experiment was shown. d, e The specificity of CD47/CD20 BsAb towards CD47 (d) or CD20 (e) expressed on Raji cells. Raji cells was co-incubated with HuNb1-biotin or Rituximab-biotin as well as Negative Ab, Rituximab (CD20 mAb), HuNb1-IgG4 (CD47 Nb) or CD47/CD20 BsAb. The binding activity was detected by FACS. f The MFI percentage of Raji cells and RBCs. Raji cells and tenfold excess RBCs were incubated with HuNb1-IgG4 or CD47/CD20 BsAb respectively. FITC positive antibodies binding to cells were detected by FACS. The RBCs were derived from three donors. g CD47/CD20 BsAb induces phagocytosis of target cells. Phagocytosis of CFSE-labeled Raji cells mediated by macrophages was analyzed by FACS. The above experiments were performed in triplicate and data were presented as mean ± SD. **P < 0.01, ***P < 0.001 versus control. h The male NOG mice were injected through tail vein with Raji cells and treated with HuNb1-IgG4, Rituximab or CD47/CD20 BsAb at the same dose (20 mg/kg), with PBS as the control (n = 6). Tumor volumes and the index of tumor volume inhibition (TGI) were shown. The experiment was performed in triplicate and one representative experiment was shown. Data were presented as mean ± SEM. **P < 0.01, ***P < 0.001 versus control