Towards Predicting Partitioning of Enzymes between Macromolecular Phases: Effects of Polydispersity on the Phase Behavior of Nonadditive Hard Spheres in Solution.

The ability to separate enzymes, or cells or viruses, from a mixture is important and can be realized by the incorporation of the mixture into a macromolecular solution. This incorporation may lead to a spontaneous phase separation, with one phase containing the majority of one of the species of interest. Inspired by this phenomenon, we studied the theoretical phase behavior of a model system composed of an asymmetric binary mixture of hard spheres, of which the smaller component was monodisperse and the larger component was polydisperse. The interactions were modeled in terms of the second virial coefficient and could be additive hard sphere (HS) or nonadditive hard sphere (NAHS) interactions. The polydisperse component was subdivided into two subcomponents and had an average size ten or three times the size of the monodisperse component. We gave the set of equations that defined the phase diagram for mixtures with more than two components in a solvent. We calculated the theoretical liquid-liquid phase separation boundary for the two-phase separation (the binodal) and three-phase separation, the plait point, and the spinodal. We varied the distribution of the polydisperse component in skewness and polydispersity, and we varied the nonadditivity between the subcomponents as well as between the main components. We compared the phase behavior of the polydisperse mixtures with binary monodisperse mixtures for the same average size and binary monodisperse mixtures for the same effective interaction. We found that when the compatibility between the polydisperse subcomponents decreased, the three-phase separation became possible. The shape and position of the phase boundary was dependent on the nonadditivity between the subcomponents as well as their size distribution. We conclude that it is the phase enriched in the polydisperse component that demixes into an additional phase when the incompatibility between the subcomponents increases.