Protein partitioning equilibrium between the aqueous poly(ethylene glycol) and salt phases and the solid protein...

True partitioning behaviour, which is independent of the protein concentration in aqueous two-phase systems, only occurs at relatively low protein concentration. The actual concentration limit depends on the properties of the protein. When the concentration of a protein exceeds relatively low values, precipitation at the interface can be observed. This protein precipitate is in equilibrium with the protein solubilized in each of the phases. This paper discusses the effect of protein solubility in view of the equilibrium of the protein concentration between the aqueous poly(ethylene glycol) and salt phases and the solid protein phase using three proteins. It was found that only rarely will the proteins be completely in solution as the concentration is increased until a solubility limit is reached and then the protein precipitates fully out of solution. A behaviour that came close to this was only seen in one case out of six. In virtually all cases, a third phase is formed which represents a solid aggregate phase which is in equilibrium with the other two, largely aqueous, phases. As the overall concentration of protein in the system is increased and the concentration in the top and bottom aqueous phases increases, the pseudo concentration in the solid-phase, C's, also increases. This could have interesting implications in terms of the amount of water associated with this phase and it certainly means that in this particular case, the solid phase is not a crystal.