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The P2A5 VHH binds to human and mouse B7-H3. SPR multi-cycle kinetic analysis of P2A5 binding to the 4-Ig (A) and 2-Ig (B) human B7-H3 isoforms, and to mouse B7-H3 (C). D, Binding kinetic parameters of P2A5 binding to the B7-H3 variants in A–C. E, Representative fluorescence microscopy images of P2A5 binding to human and mouse B7-H3. B7-H3–negative murine CT26 cells and human MKN7 cells knocked out for endogenous B7-H3 were transfected to express either murine B7-H3 or human 4-Ig B7-H3 and stained with positive control anti-mouse or anti-human B7-H3 antibodies, P2A5 as a VHH-Fc fusion molecule, or isotype control. GFP expression was used as an indicator of successful transfection.
Yeo et al. (Tue,) studied this question.