Fig. 6: Interactions between CDR H3 residues required for higher binding affinity.
The SPR sensorgrams of TREM2 binding of wild-type and variant hu3.10C2 antibodies mutated in the CDR H3 region to the corresponding residues of antibody Para.09 are shown. The sensorgrams of the parental hu3.10C2 (a), G106hD (b), D116hE (c), I107h insertion (d) single mutant, G106hD/D116hE (e), G106hD/I107h insertion (f), I107h insertion/D116hE (g) double mutant and G106hD/I107h insertion/D116hE triple mutant (h) antibodies are shown. Each panel shows the CDR H3 sequences of each variant with mutations from hu3.10C2 highlighted in red. Panels with antibodies differing by a single residue are linked by arrows from left to right-side variant, with mutations reducing or increasing binding kinetics off-rate mutation shown in green or red, respectively, and mutations with neutral or undetermined effects shown in black or gray. The binding kinetics off-rate (kd) is shown with the fold variation of two repeat experiments shown within brackets. Panels with low-quality off-rate curve fittings are indicated as “weak binding” and the effects of mutations on binding for these variants were visually assessed. Black lines in sensorgrams indicate actual data readings. Curve fittings are shown in colors, corresponding to the same antibody concentrations in different panels. RU, response units.
