Figure 1.

Thermal denaturation of V_L‐DF followed by Trp‐fluorescence and Circular Dichroism (CD). Black, scFv‐h3D6, Red, V _L‐DF. (A) Intensity of the fluorescence emission spectrum at 338 nm upon increasing temperature shows that the transition is advanced 15°C for the mutant. (B) Far‐UV CD spectra for the native states (25°C) of scFv‐h3D6 and V _L‐DF mutant show differences in the V _L signature (230 nm), whereas for the thermal‐denatured states (85°C) scFv‐h3D6 is in a β‐conformation and V _L‐DF mutant displays a singular spectrum (dotted lines). (C) Ellipticity signal at 218 nm upon increasing temperature shows that the mutant partially unfolds. Data for scFv‐h3D6 are already published and are shown for comparative purposes.13, 16