Protein folding is a complex process that is crucial for the proper functioning of biological systems. The three-dimensional structure of a protein determines its function, and misfolding can lead to various diseases, including neurodegenerative disorders. Understanding the mechanisms of protein folding and misfolding is therefore of great importance in both basic research and drug development. In this review, we provide an overview of the insights gained from biochemical studies into protein folding and misfolding. We discuss the energetics and kinetics of protein folding, the role of chaperones and other protein-folding factors, as well as the factors that contribute to protein misfolding and aggregation. Furthermore, we highlight the techniques and experimental approaches used to study protein folding and misfolding, including spectroscopic methods, mass spectrometry, and structural biology techniques. Finally, we discuss the implications of these findings for understanding disease mechanisms and for the development of therapeutic strategies targeting protein misfolding disorders.