Polymer‐based monolithic microcolumns for hydrophobic interaction chromatography of proteins

Abstract Monolithic capillary columns for hydrophobic interaction chromatography (HIC) have been prepared by thermally initiated, single‐step in situ polymerization of mixtures of monovinyl monomers including butyl methacrylate and/or 2‐hydroxyethyl methacrylate, with a divinyl crosslinker glycerol dimethacrylate or 1,4‐butanediol dimethacrylate using two different porogen systems. Two porogenic solvent mixtures were used; one “hydrophilic”, consisting of water, butanediol, and propanol, and one “hydrophobic,” comprising dodecanol and cyclohexanol. The porous structures of the monoliths were characterized and their performance was demonstrated with a separation of a mixture of myoglobin, ribonuclease A, and lysozyme under conditions typical of HIC.