Figure 4.

Analysis of anti-PD-L1 mAb binding characteristics supports 11B12-1 as a therapeutic antibody. (A) Molecular weight analysis of 11B12-1 by SDS-PAGE where M₁ indicates protein ladder, R: reducing conditions and NR: non-reducing conditions. Purified 11B12-1 is indicated by the arrow under non-reducing conditions (~100kDa). (B) Sensorgrams of binding affinities of 11B12-1 and 12F1-1 with PD-L1 analyzed by surface plasmon resonance binding assay. (C) Snapshots of antibody-antigen complex generated by AF2/ColabFold. For each antibody 20 models from 4 independent runs are overlaid. 11B12-1 is shown in red, 12F1-1 in blue, PD-L1 is depicted in green color. All structures are aligned with respect to PD-L1 coordinates. (D) Upper: 11B12-1 clashes with ligand PD-1 (ice-blue). Lower: 12F1-1 clashes with PD-L1 dimer (lime & green). Steric clashes (0.3 nm cutoff) are marked as red surface. Antibody V_L chain is shown in orange and V_H chain in cyan. (E) Immunohistochemistry staining of Syrian hamster organs with 11B12-1 and 12F1-1. Antibody clone 11B12-1 is expanded into higher magnification (20X) for each hamster organ, whereas 12F1-1 was not because of non-specific staining.