Influence of the roughness, topography, and physicochemical properties of chemically modified surfaces on the heterogeneous nucleation of protein crystals

In this study, the influence of some factors on the heterogeneous nucleation of hen egg-white lysozyme (E.C. 3.2.1.17) on a series of chemically modified surfaces has been investigated. Microbatch crystallization experiments were conducted on the microscope glass slides that were treated with poly-L-glutamic acid (PLG), poly(2-hydroxyethyl methacrylate) (P2HEMA), poly(methyl methacrylate) (PMMA), poly(4-vinyl pyridine) (P4VP), and (3-aminopropyl)triethoxysilane (APTES). An optical microscope with a heating/cooling stage was employed to measure the induction time of heterogeneous nucleation. The surface topography and roughness were characterized by atomic force microscopy. Contact angles for crystallization solution on the investigated surfaces were measured by a contact angle meter. From the theoretical analysis, the energetic barrier to heterogeneous nucleation was found to increase at higher contact angles and to decrease at higher roughness. Experimentally, a qualitative increase of the induction time of the heterogeneous nucleation on P2HEMA, APTES, and PMMA surfaces with the contact angle was observed. Such surfaces as P2HEMA, PLG, and APTES, which were of higher roughness, were shown to promote the heterogeneous nucleation. In addition, the surface with specific topography is expected to increase the possibility of the formation of a critical nucleus. Finally, the P4VP surface appeared to suppress the heterogeneous nucleation as a result of the electrostatic interaction between the lysozyme and P4VP molecules.