Thermodynamics and Single-Site Interactions between Human Serum Albumin and Green Low-Melting Mixture Solvents

Thermodynamics and interaction between human serum albumin (HSA) and low-melting mixture solvents (LoMMSs) play a critical role in the green and low-cost separation of proteins. To the best of our knowledge, there have been no reports on this issue to date. Here, we present the thermodynamic properties and single-site interaction between biobased LoMMSs betaine:glycerol and HSA. LoMMSs demonstrate strong binding affinity toward HSA, driven primarily by van der Waals forces and hydrogen bonding. LoMMSs can stabilize the secondary structure of HSA, preserving its functional integrity. This work provides valuable insights to design green, sustainable, and cost-effective LoMMSs for protein purification.