Technology trends in antibody purification.

Polymethacrylate monoliths for preparative and industrial separation of biomolecular assemblies.

Affinity monolith chromatography (AMC) is a type of liquid chromatography that uses a monolithic support and a biologically related binding agent as a stationary phase. AMC is a powerful method for the selective separation, analysis, or study of specific target compounds in a sample. This review discusses the basic principles of AMC and recent developments and applications of this method, with particular emphasis being given to work that has appeared in the last 5 years. Various materials that have been used to prepare columns for AMC are examined, including organic monoliths, silica monoliths, agarose monoliths, and cryogels. These supports have been used in AMC for formats that have ranged from traditional columns to disks, microcolumns, and capillaries. Many binding agents have also been employed in AMC, such as antibodies, enzymes, proteins, lectins, immobilized metal ions, and dyes. Some applications that have been reported with these binding agents in AMC are bioaffinity chromatography, immunoaffinity chromatography or immunoextraction, immobilized-metal-ion affinity chromatography, dye–ligand affinity chromatography, chiral separations, and biointeraction studies. Examples are presented from fields that include analytical chemistry, pharmaceutical analysis, clinical testing, and biotechnology. Current trends and possible directions in AMC are also discussed.

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Affinity monolith chromatography: a review of principles and recent analytical applications

Applications of polymethacrylate-based monoliths in high-performance liquid chromatography.

Review of recent advances in the preparation of organic polymer monoliths for liquid chromatography of large molecules.

Characterisation of grafted weak anion-exchange methacrylate monoliths

Characterization of ion exchange stationary phases via pH transition profiles.

Chemical and chromatographic stability of methacrylate-based monolithic columns

Characterisation of metal-chelate methacrylate monoliths.

Chromatographic monoliths exhibit several properties like flow-unaffected resolution and dynamic binding capacity, low pressure drop and high capacity for extremely large molecules, which are advantageous to the purification of large biomolecules such as proteins, DNA or even viruses. Recently, large-volume monolithic columns were introduced enabling the monoliths to be incorporated into industrial downstream processes. Due to the monolithic structure, however, conventional methods for resin characterization cannot be applied to monoliths, which limit their usage under cGMP conditions. In this article, a methodology of how to overcome this problem and to determine the most important parameters is presented. Several fast and noninvasive methods for monolith characterization are introduced. Their usage enables accurate determination of monolith pore size distribution and specific surface area, uniformity of the monolith as well as type and density of the active groups before or during monolith usage.

Vida Frankovič

Papers

Characterisation of grafted weak anion-exchange methacrylate monoliths

Characterization of ion exchange stationary phases via pH transition profiles.

Chemical and chromatographic stability of methacrylate-based monolithic columns

Characterisation of metal-chelate methacrylate monoliths.

Noninvasive Methods for Characterization of Large‐Volume Monolithic Chromatographic Columns