title: Comparison between protein-polyethylene glycol (PEG) interactions and the effect of PEG on protein-protein interactions using the liquid-liquid phase transition.
abstract: Phase transitions of protein aqueous solutions are important for protein crystallization and biomaterials science in general. One source of thermodynamic complexity in protein solutions and their phase transitions is the required presence of additives such as polyethylene glycol (PEG). To investigate the effects of PEG on the thermodynamic behavior of protein solutions, we report measurements on the liquid-liquid phase separation (LLPS) of aqueous bovine serum albumin (BSA) in the presence of relatively small amounts of PEG with an average molecular weight of 1450 g/mol (PEG1450) as a model system. We experimentally characterize two thermodynamically independent properties of the phase boundary: (1) the effect of PEG1450 concentration on the LLPS temperature, (2) BSA/PEG1450 partitioning in the two liquid coexisting phases. We then use a thermodynamic perturbation theory to relate the first property to the effect of PEG concentration on protein-protein interactions and the second property to protein-PEG interactions. As criteria to determine the accuracy of a microscopic model, we examine the model's ability to describe both experimental thermodynamic properties. We believe that the parallel examination of these two properties is a valuable tool for verifying the validity of existing models and for developing more accurate ones. For our system, we have found that a depletion-interaction model satisfactorily explains both protein-PEG interactions and the effect of PEG concentration on protein-protein interactions. Finally, due to the general importance of LLPS, we will experimentally show that protein-PEG-buffer mixtures can exhibit two distinct types of liquid-liquid phase transitions.
authors.affiliation: Department of Chemistry, Texas Christian University, Fort Worth, Texas 76129, USA.
authors.forename: Ying
authors.initials: Y
authors.lastname: Wang
authors.forename: Onofrio
authors.initials: O
authors.lastname: Annunziata
mesh_terms: D044367:Phase Transition; D011092:Polyethylene Glycols / Q000737:chemistry*; D011485:Protein Binding; D012710:Serum Albumin, Bovine / Q000737:chemistry*; D012996:Solutions / Q000737:chemistry; D013816:Thermodynamics; D014867:Water / Q000737:chemistry
chemical_list: D012996:Solutions; D014867:Water; D012710:Serum Albumin, Bovine; D011092:Polyethylene Glycols