Figure 1
From: An engineered antibody binds a distinct epitope and is a potent inhibitor of murine and human VISTA
Engineering a cross-reactive anti-VISTA antibody. (a) Library screening progression used to isolate scFv variants that bound human VISTA (Round 1) and mouse VISTA (Round 2). Flow cytometry gates used for screening are shown on dot plots of individual sorts. X-axis depicts expression of scFv on the yeast cell surface, y-axis depicts VISTA binding, measured by antibodies against c-myc and VISTA, respectively. (b) Binding intensity to human VISTA-Fc (red) and mouse VISTA-His (gray) of individual scFv clones isolated after Round 1 of screening. The V9 clone displayed above background binding signal to mouse VISTA-His. (c) Sequence comparison of scFv clones after the first round (V9, V11, V13, V15) and after the second round (V9.3 and V9.7) of sorting. Amino acid differences from final V9.7 clone are highlighted in red. (d) Full kinetic binding curves of SG7 (antibody form of top clone V9.7) against human VISTA-His and mouse VISTA-His monomers, as measured by KinExA. Mean ± standard deviation of duplicate measurements are shown for D.
