Room temperature stable agarose solutions

Abstract

A method for making functionalized porous crosslinked polysaccharide gel coated structures used as liquid chromatography media is provided. The method includes impregnating a porous substrate with a room temperature stable aqueous polysaccharide solution containing water, 0.1% to 20% of a polysaccharide, 18% to 54% of a gel-inhibiting agent to prevent the gel from re-gelling, and 0.001% to 10% of an anionic fluorosurfactant for optimum solution coatability onto the substrate, each concentration is by total weight of the aqueous solution. Next water is evaporated from the coating, followed by exposing the dehydrated coating to a gelling agent thereby forming a porous polysaccharide gel coated substrate. Next the gel coated substrate is exposed to a crosslinking agent forming a porous crosslinked polysaccharide gel coated substrate. Next, the gel coated substrate is functionalized by contacting the porous polysaccharide gel coated substrate with sodium 3-bromopropanesulfonate, thereby attaching sulfopropyl ligands to the gel coated substrate, resulting in a functionalized porous absorptive polysaccharide gel coated chromatography structure. When the functionalized gel coated chromatography structure is used as liquid chromatography media, and a protein (e.g. lysozyme) containing solution contacts the media, the sulfopropyl ligands attached to the gel coated chromatography structure will bond to the protein, thereby removing the protein from the protein containg solution.