Fabrication and characterization of superporous cellulose bead for high-speed protein chromatography

Novel superporous cellulose (SC) matrix has been fabricated by water-in-oil emulsification-thermal regeneration using granules of calcium carbonate as porogenic agents. As a control, microporous cellulose (MC) bead was fabricated in the absence of calcium carbonate. Simultaneously, double cross-linking was applied to enhance the mechanical strength of the particles. The photographs by scanning electron microscopy of the SC bead illustrated that there were more "craters" of several microns scattering on the surface of the beads. It led to a higher water content and effective porosity of the SC medium. The two beads were then modified with diethylaminoethyl (DEAE) group to prepare anion exchangers. The dynamic uptake results of bovine serum albumin (BSA) exhibited that the pore diffusivity of BSA in the DEAE-SC bead was two to three times larger than that in the DEAE-MC bead. In addition, the column packed with the DEAE-SC showed lower backpressure, higher column efficiency and dynamic binding capacity than the column packed with the DEAE-MC at a flow rate range of 150-900cm/h. Moreover, the column efficiency of the DEAE-SC column was independent of flow velocity up to a flow rate of 1200cm/h. All the results exhibited the superior characteristics of the SC bead as a potential medium for high-speed protein chromatography.