Figure 4

Characterization of the ΔT_m shift. (A) Concentration dependence of apparent T_m values measured for four nanobodies and plotted in a zero-centered fashion around the T_m values at 12.7 µM; orange curve: nanobody with a second disulfide bond; blue curves: nanobodies devoid of a second disulfide bond. Vertical dashed lines indicate the concentrations used in the experiments described before. Error bars represent the average standard error of 0.13 °C for a T_m measurement. (B) Histrogram of ΔT_m values calculated for 57 nanobodies using the equation ΔT_m = T_m(13.1 µM) − T_m(32.7 µM). 11 nanobodies were excluded since at one of the two concentrations a T_m value could not be unambiguously assigned.