(a) SPR kinetic analysis of RH5 binding with twofold dilutions of RH5 from a maximal concentration of 8 μM and threefold dilutions of RH5 from a maximal concentration of 333 nM for BSGdes7 (kinetic fits shown in gray; ka=6.3*104 M-1s-1, kd = 0.14 s-1, KD=2.2 μM, in agreement with ref. 10,14) and ka=5.6*104 M-1s-1, kd = 6.4*10-5 s-1
KD=1.15 nM for BSG and BSGdes7, respectively. (b) BSGdes7 is predicted to exhibit improved core packing and reduced electrostatic strain at the core of the interface. (c) Thermal denaturation of BSG and BSGdes7.