Use of Ceramic Monoliths as StationaryPhase in Affinity Chromatography

Abstract

The use of coated ceramic monoliths as support for affinity chromatography is described. Ceramic monoliths are robust active matrix supports and present a very small pressure drop. Monoliths are coated with a very thin agarose gel layer and activated using a standard activation process for agarose beads. Experiments demonstrate that enzyme adsorption occurs exclusively on the outside surface of the agarose coating since enzyme molecules are too large to fit into the porous matrix. Adsorption and desorption rates are large and production of enzyme per unit monolith volume justifies further exploring this separation process for large throughput operation.

Monoliths for fast bioseparation and bioconversion and their applications in biotechnology

“…Examples of monoliths for bioseparation and biotransformation are polymethacrylate block polymers (CIM disksm, CIM tubesm, Swiftm columns) produced directly by free radical polymerization induced thermally (for reviews see [2 -8]) or by radiation [9], polymethacrylate based monoliths with templated pores [10,11], monoliths prepared from compressed polyacrylamide gels [12 -14], silica columns manufactured as single blocks by a sol-gel process [15 -17], silica xerogels [18], emulsion-derived polymeric foams such as polyHIPEm [19], monoliths prepared via metathesis polymerization [20], continuous urea-formaldehyde resins [21], monoliths prepared from carbon microspheres [22,23], graphitized carbon monoliths [22], monoliths cast from cellulose [24] and superporous agarose [25], cryogels from polyacrylamide [26 -28] and poly(vinyl alcohol) [29 -31], polyacrylamide-coated ceramics [32], rolled woven fabrics [33,34], and adsorptive membranes of various types [35 -46]. As long as they are used as a single sheet, membranes can be considered as monoliths with an extreme geometry in which the dimension in the axial direction is very short.…”

Bioaffinity chromatography on monolithic supports

“…Bioaffinity applications concerning high-resolution separations of closely related compounds, e.g. chiral separations [29][30][31][32][33][34] are not discussed. The emphasis is on selective retention followed by nonspecific or biospecific elution, i.e.…”

The suitability of DEAE-Cl active groups on customized poly(GMA-co-EDMA) continuous stationary phase for fast enzyme-free isolation of plasmid DNA

“…There exists generally a trade-off between pressure drop and binding capacity as increasing pore size decreases binding capacity (decreasing surface area) and decreasing pore size increases pressure drop. Different types of monolithic supports currently available are cryogels from polyacrylamide [6,7], emulsion-derived monoliths [8], polymethacrylate based polymers synthesised by free radical polymerisation induced thermally [8][9][10][11][12][13] or by radiation [14], silica columns manufactured as single blocks by a sol-gel process [15,16], silica xerogels [17], monoliths prepared from compressed polyacrylamide gels [18,19], polymer monoliths prepared through metathesis [20], monoliths prepared from carbon microspheres [21,22], carbon monoliths [21], cellulose-based monoliths [23], superporous agarose gel [24], poly vinyl alcohol [25], polyacrylamide-coated ceramics [26] and rolled woven fabrics [27]. Polymethacrylate monolithic supports are optimal adsorbents for pDNA separation.…”