posted on 2024-01-27, 01:04authored byJi Young Yang, Oliver Burkert, Boris Mizaikoff, Jens Smiatek
We performed nano
differential scanning fluorimetry (nanoDSF) measurements
of immunoglobulin G (IgG) in urea gradient solutions under thermal
unfolding. Our results show that the denaturing effect of urea on
individual IgG domains can be monitored via a linear mapping of thermal
shift curves to the corresponding urea concentrations. Assignment
of IgG domains to each thermal shift curve allows for a reliable differentiation
of the underlying mechanisms. Further results show a decisive influence
of salt-induced electrostatic screening effects. We are able to explain
all findings by preferential binding mechanisms in combination with
electrostatic effects. The results of our study shed more light on
the complex interaction mechanisms between buffer solutions and complex
proteins, which are important for improving the shelf life of protein
therapeutic formulation.