The kinetics of homogeneous and two‐step nucleation during protein crystal growth from solution

Many experimental reports for the kinetics of crystal nucleation and growth, from an isothermal solution, point to a sigmoidal‐like behavior for the process. Here we consider three different nucleation models from the literature and show that all lead to sigmoidal or sigmoidal‐like behavior for the kinetics of nucleation. A two‐step nucleation process is known to occur within certain supersaturated protein solutions, and it is demonstrated in this report how the sigmoidal law yields kinetic information for the two‐step and homogeneous nucleation modes. We propose here that two‐step solute‐rich associates form in the solution around seed nuclei that are already present at or near the point in time when the solution is prepared. Using this hypothesis, we are able to model the time‐dependent volume of the two‐step phase per unit volume of solution and show that this compares well with reported experimental data. A kinetic model is given for the proposed process, which leads to a sigmoidal rate law. Additionally, a relation between the initial and final nuclei densities and the induction time is derived. As a result of this study, the conclusion is that two‐step activity increases with increasing initial supersaturation or increasing salt concentration.