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This lecture delves into the intricate process of protein folding, exploring the key forces involved such as van der Waals interactions, the hydrophobic effect, hydrogen bonds, and salt bridges. It also discusses the destabilizing factors like chain entropy and bond strain, providing insights into the complex interplay of stabilizing and destabilizing forces. The lecture further covers the estimation of net magnitudes of these forces in protein folding and the thermal melting of proteins. Additionally, it examines the applications of ThermoFluor in studying protein stability and ligand binding, shedding light on how ligand binding can stabilize protein structures.