Hydrophilization of gigaporous polystyrene microspheres with saccharide as high-speed protein chromatography base support

Gigaporous poly(styrene-divinylbenzene) (PS) microspheres were hydrophilically modified with natural saccharide to minimize their nonspecific adsorption to proteins. The microspheres were chloroacetylated through Friedel-Crafts acetylation with chloroacetyl chloride, and then coupled with diacetone-D-glucose (DAGlu) through the Williamson reaction, and the protecting groups were removed on DAGlu. Results showed that the PS microspheres were successfully coupled with DAGlu and that the gigaporous structure was well maintained. After hydrophilization (Glu-PS), nonspecific adsorption of proteins on PS microspheres was greatly reduced. The high surface density of hydroxyl groups on Glu-PS microspheres surface make it easy to derivatize the spheres by classical methods. Flow experiments showed that the Glu-PS column had low backpressure, good permeability, and mechanical stability. All results indicate that the Glu-PS microspheres have great potential applications in high-speed protein chromatography.