Ostwald-like ripening of the anomalous mesoscopic clusters in protein solutions

Metastable clusters of mesoscopic dimensions composed of protein-rich liquid exist in protein solutions, both in the homogeneous region of the solution phase diagram and in the region supersaturated with respect to an ordered solid phase, such as crystals; in the latter region they are crucial nucleation sites for ordered solids. We monitor, using three optical techniques, the long-term evolution of the clusters in lysozyme solutions at conditions where no condensed phases, liquid or solid, are stable or present as long-lived metastable domains. We show that cluster formation is a reversible process and that the clusters are in near equilibrium with the solution, up to a capillary correction. In contrast to classical phase transformations, the solution concentration at cluster-solution equilibrium is close to its initial value; this is akin to chemical reaction equilibria and demonstrates the complex chemical composition of the clusters. However, similar to classical phase transformations, en route to full equilibration, the average cluster size grows with time following a universal law t(0.26±0.03), independent of the cluster volume fraction; the cluster size distribution is scale-invariant at all stages of cluster evolution. Despite the correspondence of these behaviors to the Lifshitz-Slyozov-Wagner (LSW) theory predictions, the cluster sizes are about 10× smaller than the LSW prediction, likely due to the complex cluster composition. The observed cluster evolution helps us to understand nucleation mysteries, such as nucleation rates lower by orders of magnitude than classical theory predictions, nucleation rate variable under steady conditions, and others.