Polymer coatings for improved protein crystal growth

The ability to grow quality protein crystals is necessary to analyze protein structure by X-ray diffraction and related techniques. As such it plays a key role in enzymology, structure-based drug design, molecular biology, and other biomedical areas. It is also required for macromolecule purification by crystallization. Protein crystal growth (PCG) may be negatively influenced by various factors related to nonspecific adsorption and adherence at growth chamber surfaces. Such factors include nucleation and growth of flawed crystals at chamber walls, or wall growth blockage of optical monitoring paths. Surface localized poly(ethylene glycol) (PEG) and other neutral, hydrophilic polymers are known to significantly reduce nonspecific adsorption of biological macromolecules and particles. Preliminary studies, involving various PCG methods (temperature induction, vapor diffusion), apparatii (test tubes, cuvettes, and specialized PCG hardware), growth chamber materials (glass, polystyrene, polysulfone), chamber volumes (0.1–10 ml) and protein samples (lysozyme, thaumatin, insulin) indicate the potential of PEG coatings to significantly reduce problems related to adsorption in PCG. The results, which match the ability of such coatings to reduce protein adsorption as evaluated by both ellipsometry and enzyme linked immunoassay, are discussed in relation to colloidal stabilization theory and properties of PEG coated surfaces.