Search

A novel super-macroporous monolithic composite cryogel was prepared by embedding macroporous cellulose beads into poly(hydroxyethyl methacrylate) cryogel. The cellulose beads were fabricated by using a microchannel liquid-flow focusing and cryopolymerization method, while the composite cryogel was prepared by cryogenic radical polymerization of the hydroxyethyl methacrylate monomer with poly(ethylene glycol) diacrylate as cross-linker together with the cellulose beads. After graft polymerization with (vinylbenzyl)trimethylammonium chloride, the composite cryogel was applied to separate immunoglobulin-G and albumin from human serum. Immunoglobulin-G with a mean purity of 83.2% and albumin with a purity of 98% were obtained, indicating the composite cryogel as a promising chromatographic medium in bioseparation for the isolation of important bioactive proteins like immunoglobulins and albumins., (© 2013 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim.)

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., (Copyright © 2013 Elsevier B.V. All rights reserved.)

Polymeric cryogels are sponge-like materials with supermacroporous structure, allowing them to be of interest as new chromatographic supports, cell scaffolds and drug carriers in biological and biomedical areas. The matrices of cryogels are always prepared in the form of monoliths by cryo-polymerization under frozen conditions. However, there are limited investigations on the production of cryogels in the form of adsorbent beads suitable for bioseparation. In this work, we provide a new approach by combining the microchannel liquid-flow focusing with cryo-polymerization for the preparation of polyacrylamide-based supermacroporous cryogel beads with a narrow particle size distribution. The present method was achieved by introducing the aqueous phase solution containing monomer, cross-linker and redox initiators, and the water-immiscible organic oil phase containing surfactant simultaneously into a microchannel with a cross-shaped junction, where the aqueous drops with uniform sizes were generated by the liquid shearing and the segmentation due to the steady flow focusing of the immiscible phase streams. These liquid drops were in situ suspended into the freezing bulk oil phase for cryo-polymerization and the cryogel matrix beads were obtained by thawing after the achievement of polymerization. By grafting the polymer chains containing sulfo binding groups onto these matrix beads, the cation-exchange cryogel beads for protein separation were produced. The results showed that at the aqueous phase velocities from 0.5 to 2.0 cm/s and the total velocities of the water-immiscible phase from 2.0 to 6.0 cm/s, the obtained cryogel beads by the present method have narrow size distributions with most of the bead diameters in the range from 800 to 1500 μm with supermacropores in sizes of about 3-50 μm. These beads also have high porosities with the averaged maximum porosity of 96.9% and the mean effective porosity of 86.2%, which are close to those of the polyacrylamide-based cryogel monoliths. The packed bed using the cryogel beads with mean diameter of 1248 μm, as an example, has reasonable and acceptable liquid dispersion, but high water permeability (4.29 × 10⁻¹⁰ m²) and high bed voidage (90.2%) owing to the supermacropores within the beads, enhanced the rapid binding and separation of protein from the feedstock even at high flow velocities. The purity of the obtained lysozyme from chicken egg white by one-step chromatography using the packed bed was in the range of about 78-92% at the flow velocities of 0.5-15 cm/min, indicating that the present cryogel beads could be an effective chromatographic adsorbent for primary bioseparation., (Copyright © 2012 Elsevier B.V. All rights reserved.)

Graphical abstract Highlights • Slug flow of aqueous Dex-MA-PEGDA and ethyl acetate system in microchannels was characterized. • Slug formation mechanism was revealed by analyzing the forces acting on the two phases. • Flow regime map for the controllable fabrication of dextran-based cryogel beads was obtained. • Correlations of the slug lengths, the droplet sizes and the cryogel bead diameters were proposed. • Supermacroporous dextran-based cryogel beads with a narrow size distribution were fabricated. Abstract Cryogel beads are novel adsorbents with potential applications in biotechnology, environmental and tissue engineering. However, the controllable preparation of cryogel beads with expected small and uniform diameters is a challenging work. The micro-flow-focusing method combining with the cryo-polymerization has been suggested as a new approach for the fabrication of cryogel beads. In order to form the uniform droplet, the slug flow is generated in the microchannel, so the slug flow hydrodynamics are crucial and fundamental for this process. In the present work, the dextran-methacrylate (Dex-MA-PEGDA) solution and ethyl acetate containing surfactant of Span 80 were used as the aqueous phase and the water-immiscible, respectively. The fabrication of dextran-based cryogel beads and the slug flow hydrodynamics in a rectangular microchannel were investigated experimentally. The flow regime map was proposed and the typical regime boundaries were measured and correlated, and the formation mechanisms of slugs were also discussed. The regime boundaries were correlated by Weber number and Capillary number, i.e., We o = 58.16 Ca w 0.92 for the boundary between the annular and the slug flow, and We o = 4.12 Ca w 0.09 for the slug and the bubbly flow, respectively. Several crucial parameters of the droplets generated by the slug flow, i.e., the aqueous slug length, the aqueous slug velocity and the aqueous droplet size, were measured and correlated. The Dex-MA-PEGDA cryogel beads were prepared successfully under different conditions and their diameters were correlated to the sizes of the droplets. Eventually, the uniform size distributions and supermacroporous microstructures of dextran-based cryogel beads were demonstrated. [ABSTRACT FROM AUTHOR]