Poly(glycidyl methacrylate-divinylbenzene-triallylisocyanurate) continuous-bed protein chromatography

A novel continuous bed with high dynamic adsorption capacity for protein has been developed. It is a macroporous poly(glycidyl methacrylate-divinylbenzene-triallylisocyanurate) rod prepared by in situ copolymerization in a chromatographic tube. The bed matrix contained epoxy groups, so diethylaminohydroxypropyl groups were coupled to the matrix, leading to an anion-exchange continuous bed. The component, specific surface area, and the pore structure of the bed matrix were characterized by Fourier transform infrared spectroscopy, BET method and scanning and transmission electron microscopies, respectively. The flow properties, column efficiency and the dynamic adsorption behavior of the bed were studied. The results show that the continuous bed, a ternary copolymer of glycidyl methacrylate (GMA), divinylbenzene (DVB) and triallylisocyanurate (TAIC) with a specific surface area of 56.4 m2/g, consisted of a three-dimensional structure made up of continuous clusters of microspheres (300 nm) and interconnected irregular pores. The rate of mass transfer is enhanced by the convection of the mobile phase through the pores. The dynamic adsorption isotherm of the anion-exchange column for bovine serum albumin was expressed by the Langmuir equation with a dynamic capacity as high as 76.0 mg/g. Moreover, the separation of proteins, i.e. lysozyme, hemoglobin and bovine serum albumin, is little affected by mobile-phase velocity up to 902.5 cm/h; it was completed within 5 min at 902.5 cm/h.