The aim of this diploma thesis was to experimentally investigate the thermodynamic stability and conformational changes of the SG4 nanobody. In this study, we employed and compared the results of two complementary physicochemical methods: differential scanning calorimetry (DSC) and UV-Vis spectroscopy.
Differential scanning calorimetry experiments enabled direct monitoring of the thermal denaturation of the SG4 nanobody and the determination of key thermodynamic parameters, such as the midpoint transition temperature, calorimetric enthalpy, and van 't Hoff enthalpy. Conducting multiple consecutive heating and cooling cycles served as a method to evaluate the reversibility of the conformational transition of SG4.
UV-Vis spectroscopic analysis was used as an additional independent method to study the temperature dependence of absorbance. The results obtained for the SG4 nanobody using both methods were compared regarding the assumptions of the two-state model and the physical background of the measurements themselves. The insights gained contribute to a better understanding of nanobody structural stability.
|