Figure 2.

Fraction of aggregation-free nanobodies. (A) Size-ranked turbidity integrals obtained during the heating phase. Integrals for nanobodies devoid of a significant turbidity signal were set to zero (data points to the right of the colored, vertical lines; red: 32.7 µM, orange: 13.1 µM). Data points in a single column refer to the same nanobody. Name-labeled data points correspond to the kinetically characterized nanobodies of Fig. 3A with aggregation rate constants in blue and in s⁻¹. For error calculation see Methods section; for raw traces see Supplementary Figure 1. (B) Turbidity integrals as in panel A but for the cooling phase. The percental fractions of aggregation-free nanobodies refer to the full set of 68 binders. For raw traces see Supplementary Figure 2. (C) Fraction of reversibly refolding nanobodies judged from differences between initial and final fluorescence ratios. For calculation see Methods section. Values for 32.7 (red) and 13.1 µM (orange) were separately ranked by size. Horizontal gray line: Threshold of significance (three times the average standard error observed in initial and final intervals of a 2 °C range).