Evaluation of Slow Equilibration Kinetics in Antibody Chemical Denaturation Experiments with the Prometheus NT.Plex

Chemical denaturation is becoming increasingly popular in the drug development process, allowing determination of the free enthalpy (ΔG) of unfolding of monoclonal antibodies (mAbs), which directly correlates with the fraction of unfolded protein in solution and provides thermodynamic insight into mAb stability. In this technical note, the Prometheus NT.Plex is used to measure the isothermal equilibration of chemical denaturation reactions for three different mAbs at GuaHCl concentrations between 0 and 7 M at 20 °C.