Protein crystallization by using porous glass substrate

Abstract

The effects of a commercially available porous glass substrate (Corning Porous Glass No.7930) on the heterogeneous nucleation of proteins [hen egg‐white lysozyme (HEWL), thaumatin and apoferritin] have been investigated in order to develop an improved method to facilitate the nucleation of protein crystals. It was found that the porous glass substrate could promote the nucleation at lower supersaturations. The induction time for nucleation decreased, and the crystals obtained from porous glass substrates were larger than those from normal glass substrates. Many pores and channels of 10–100 nm in diameter were observed on the porous glass surface by atomic force microscopy (AFM). It is believed that these pores and channels are crucial for facilitating the nucleation process in this work.

Trypsin crystallization by membrane-based techniques

“…As already demonstrated for HEWL in previous work (Di ProWo et al, 2003), an accelerated protein crystal growth can be obtained with the membrane-based system, due to the chemical-physical properties of the polymeric surface. In fact, in accordance with information published in literature, this eVect is imputable both to the porous structure (Chayen et al, 2001;Rong et al, 2004) and the hydrophobic nature (Fermani et al, 2001;García-Ruiz, 2003) of the polymeric surface that, acting as active support for heterogeneous nucleation, induces an acceleration of the crystallization kinetics.…”

Selective Crystallization of Proteins Using Engineered Nanonucleants

“…Indeed it is argued that the lack of selectivity exhibited by these materials is one of their primary benefits. It has been previously reported that surfaces with narrow pore size distributions and structured pore arrangement failed to influence nucleation and crystallization 16,22,23 . This could be due to inappropriate pore size range or chemistry of the substrate used.…”

Effects of forced solution flow on lysozyme crystal growth

“…Further AFM studies demonstrated that the flow changed the surface morphology drastically. For crystals grown under forced flow condition, the step density is higher and the step growth rates are lower compared with those grown under quiescent condition [18,19]. At present, there is no further explanation why the forced flow can enhance the crystal quality, and thus the influence of convection transport on the crystal growth is unclear.…”

“…Forced solution flow has been employed in protein crystallization experiments [15][16][17][18][19][20] to investigate quantitatively the effects of solution convection on protein crystal growth. Rosenberger et al designed a solution circulation loop, including a circinal cell and a peristaltic pump [13,15], and found from experiments that whether the growth rates increased depended on the flow rates and purity of lysozyme.…”