Denatured protein analysis program
This program evaluates a single protein substance in solution that is in equilibrium at a certain temperature, and enables calculation of the thermodynamic parameters (Tm, ΔH, ΔS) of its protein from the data with variable temperatures.
KD is the ratio of [N] and [D], and is calculated by using the experimental data, which is the measured CD values θi as a function of temperature Ti. This program approximates θi by the following equation, and calculates the KD.
Application
Denaturation study of the VHH antibody
Figure shows the denaturation curve of the CD at 217 nm which refl ects changes in β-sheet structure of the VHH antibody. Comparing the denaturation temperatures, it can be seen that the thermal stability of the samples decreases in the following order: No. 3, 1, and 2.
Temperature vs. CD denaturation curves
Anti-SARS-Cov-2 VHH antibody a) No. 1 (0.4 mg/mL), b) No. 2 (0.28 mg/mL), and c) No. 3 (0.2 mg/mL).