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. 2021 Jan 5;10:615534. doi: 10.3389/fonc.2020.615534

Figure 2.

Figure 2

Characterization of antihuman CD47 monoclonal antibody 2C8. (A) An antibody subtype was detected by the Mouse Monoclonal Antibody Isotyping Kit. (B) VH and VL regions amplified from hybridoma RNA/cDNA. M: marker; Lane 1: 2C8-VL region (330 bp); Lane 2: 2C8-VH region (351 bp). (C) UV spectrum of purified 2C8. (D) SDS-PAGE was used to show the purified 2C8 antibodies (M: marker, Lane 1, Mouse ascites before purification; Lane 2: purification flow through; Lane 3: purified 2C8). (E). Flow cytometry analysis shows that 2C8 shows a high affinity for CD47, which was 0.2991×10-9 M. (F) MFI demonstrates that 2C8 disrupts the binding of APC labeled B6H12.2 with Daudi cells. (G) Antibody homology modeling structure of 2C8. (H) Compound structure of B6H12.2/CD47-ECD superimposed on 2C8/CD47-ECD showing a shared molecular docking. Residues were interacting with only B6H12.2 (blue), only 2C8 (red), or both ligands (green).