Crystallization of Proteins at Ultralow Supersaturations Using Novel Three-Dimensional Nanotemplates

A series of novel three-dimensional (3D) nanotemplates which have tuned surface mesoporosity and surface chemistry based on the protein of interest have been developed to facilitate protein crystallization. The crystallization of five model proteins systems is reported at hereto the lowest reported protein or precipitant concentrations. These improvements were only possible due to the combined use of optimum pore sizes with appropriate surface chemistries in the preparation of the 3D nanotemplates. The success of this strategy can be ascribed to the specific design of the ordered nanotemplates which are based on known physicochemical properties of the protein and offer an alternate targeted strategy for protein crystallization in contrast to previous methods based on the use of universal nucleants. The use of protein tuned nanotemplates will potentially open up new opportunities for the crystallization and structure determination of high value proteins, as well as opportunities for their separation and purification in downstream bioprocessing.