Polymethacrylate‐based monoliths as stationary phases for separation of biopolymers and immobilization of enzymes

Abstract

The experiences in the production and application of polymethacrylate-based monolithic supports, since their development almost thirty years ago, are presented. The main driving force for the development of new chromatographic supports was the necessity for the isolation and separation of physiologically active biopolymers and their use for therapeutic purposes. For this sake, a development of a method for fast separation, preventing denaturation and preserving their biological activity was necessary. Development of polysaccharide-based supports, followed by the introduction of polymer-based chromatographic media, is shortly described. This development was followed by the advances in monolithic media that are now used for both large- and small-scale separation of biopolymers and nanoparticles. Finally, a short overview is given about the applications of monoliths for sample displacement chromatography, resulting in isolation of physiologically active biomolecules, such as proteins, protein complexes, and nucleic acid, as well as high-throughput sample preparation for proteomic investigations.

Molecularly imprinted macroporous polymer monolithic layers for L‐phenylalanine recognition in complex biological fluids

“…20,21 In contrast to the diffusively controlled bead-based columns, the monoliths are characterized by convective mechanism of mass transfer. Besides the wide application of flow-through monoliths for various dynamic processes, [22][23][24] these materials have been also found to be highly prospective in a view of their application as 3D biochip platforms (microarrays) for analysis of both proteins 25,26 and DNA. 27,28 At present time, there is a range of publications devoted to development and application of macroporous MIP monoliths in flow-through format for isolation and separation of various small molecules including drugs, pesticides, herbicides, amino acids, and so on.…”

Flow‐through enzymatic reactors using polymer monoliths: From motivation to application

“…The first generation of polymer monoliths developed in 1970s suffered the problem of excessive swelling, even softening in some solvents, which seriously limited their application [28]. In 1990s, Svec and Fréchet developed rigid polymer monoliths using “molding” approach, that is, in situ polymerization of organic monomers and crosslinkers in the presence of porogenic solvents [29,30].…”

Sample preparation in foodomic analyses

“…However, each separation or enrichment during sample preparation produces large number of fractions. Final result (and price) is significant reduction of sample throughput, introduction of experimental variability, and in an increase in the costs of the analysis . A widely used method is filter‐aided sample preparation, recommended by Wiśniewski .…”

“…Routine application of MALDI MS in food analysis, especially for fast identification of bacterial contaminations and food adulterations , requests further miniaturization, and introduction of on‐chip sample preparation as well as reduction of the sample amount . These strategies are further driving forces toward the introduction of high‐throughput (not only proteomic, but also other foodomic) methods, use of laboratory robotics, and introduction of special chromatographic media such as monolithic supports and membranes , as well as magnetic beads , toward analyses of a large number of samples .…”