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    From adsorption to crystallization of proteins: Evidence for interface-assisted nucleation

    Year: 2024

    Authors: Banks, Hadra; Surfaro, Furio; Pastryk, Kai-Florian; Buchholz, Cara; Zaluzhnyy, Ivan A.; Gerlach, Alexander; Schreiber, Frank

    Keywords: Heterogeneous nucleation; Human serum albumin; Protein adsorption; Protein crystallization; QCM-D

    Protein crystallization is among the key processes in biomolecular research, but the underlying mechanisms are still elusive. Here, we address the role of inevitable interfaces for the nucleation process. Quartz crystal microbalance with dissipation monitoring (QCM-D) with simultaneously optical microscopy, confocal microscopy, and grazing-incidence small angle X-rays scattering (GISAXS) were employed to investigate the temporal behavior from the initial stage of protein adsorption to crystallization. Here we studied the crystallization of the Human Serum Albumin (HSA), the most abundant blood protein, in the presence of a charged surface and a trivalent salt. We found evidence for interface-assisted nucleation of crystals. The kinetic stages involved are initial adsorption followed by enhanced adsorption after longer times, subsequent nucleation, and finally crystal growth. The results highlight the importance of interfaces for protein phase behavior and in particular for nucleation.
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