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Journal of chromatography. A

Rapid freezing cryo-polymerization and microchannel liquid-flow focusing for cryogel beads: adsorbent preparation and characterization of supermacroporous bead-packed bed.


PMID 23453460

Abstract

Cryogel beads, fabricated by the microchannel liquid-flow focusing and cryo-polymerization method, have micron-scale supermacropores allowing the passage of crude feedstocks, and could be of interest as chromatographic adsorbents in bioseparation applications. In this work, we provide a rapid freezing and continuous formation method for cryogel beads by cryo-polymerization using dry ice particles as the freezing source and microchannel liquid-flow focusing using peristaltic pumps for the fluid supply. Polyacrylamide (pAAm)-based supermacroporous cryogel beads were prepared and grafted with N,N-dimethylaminoethyl methacrylate (DMAEMA), which provided the anion-exchange cryogel beads with tertiary amine functional groups suitable for binding proteins. Properties of the supermacroporous cryogel-bead packed bed, i.e., permeability, bed voidage, protein breakthrough as well as protein adsorption performance by using bovine γ-globulin as model protein, were experimentally investigated. A capillary-based model was employed to characterize the supermacroporous bed performance, and gave a reasonable description of the microstructure and thus an insight into the flow, dispersion and mass transfer behaviors within the cryogel bead-packed bed. The results also showed that by using dry ice as the freezing source, it is easy to reduce the temperature below -55 to -61°C in the bulk solution, causing the rapid formation of ice crystals within the monomer drops, and finally effective cryo-polymerization to form supermacropores within the cryogel beads. By using peristaltic pumps, continuous preparation was achieved and the obtained cryogel beads had favorable properties similar to those prepared using syringe pumps in the microchannel liquid-flow focusing process. This method is thus expected to be interesting in the liter- or even larger-scale preparation of cryogel adsorbents.