Nanodiamonds as Nucleating Agents for Protein Crystallization.

Nanodiamond (ND) is a carbon-based nanomaterial with potential for a wide range of biological applications. One of such applications is to facilitate the nucleation of protein crystals in aqueous solution. Here, we show that NDs (nominal diameters of 30 and 100 nm) after surface oxidation in air and subsequent treatment in strong acids are useful as heterogeneous nucleating agents for protein crystallization. Tested with lysozyme, ribonuclease A, proteinase K, and catalase, the nanomaterials in either aggregate or film form are found to be able to increase the crystallization efficiency of all proteins. Particularly, for 30 nm NDs, the films with an area of ∼2 mm2 can effectively induce the crystallization of lysozyme at a concentration as low as 5 mg/mL. The efficiency can be further improved by adding preformed protein clusters (∼300 nm in diameter) as inherent nucleation precursors, as demonstrated for ribonuclease A. This combined approach is easy to implement, highly compatible with existing technologies, and can be applied to other protein samples as well.