Affinity monolith chromatography: A review of general principles and recent developments

Abstract

Affinity monolith chromatography (AMC) is a liquid chromatographic technique that utilizes a monolithic support with a biological ligand or related binding agent to isolate, enrich, or detect a target analyte in a complex matrix. The target‐specific interaction exhibited by the binding agents makes AMC attractive for the separation or detection of a wide range of compounds. This article will review the basic principles of AMC and recent developments in this field. The supports used in AMC will be discussed, including organic, inorganic, hybrid, carbohydrate, and cryogel monoliths. Schemes for attaching binding agents to these monoliths will be examined as well, such as covalent immobilization, biospecific adsorption, entrapment, molecular imprinting, and coordination methods. An overview will then be given of binding agents that have recently been used in AMC, along with their applications. These applications will include bioaffinity chromatography, immunoaffinity chromatography, immobilized metal‐ion affinity chromatography, and dye‐ligand or biomimetic affinity chromatography. The use of AMC in chiral separations and biointeraction studies will also be discussed.

Matrices and Affinity Ligands for Antibody Purification and Corresponding Applications in Radiotherapy

“…Monoliths have been extensively applied in antibody purification with different affinity ligands, such as protein A [ 28 ], L-histidine [ 29 ], and peptides [ 30 , 31 ]. Further discussion on affinity monoliths has been reported [ 26 , 32 ], and this review will not repeat it.…”

Functionalized cryogel monoliths for fast and selective separation of nucleic acids directly from crude lysate

“…Post synthesis, cryogels retain their shape of the reaction pot in which they have synthesized (Figure 1). Due to the interconnectivity property of the pores, the mobile phase easily flows through convection that generates the very least flow resistance (Napolitano-Tabare et al, 2021;Poddar et al, 2021;Tao et al, 2015).…”

“…Post synthesis, cryogels retain their shape of the reaction pot in which they have synthesized (Figure 1). Due to the interconnectivity property of the pores, the mobile phase easily flows through convection that generates the very least flow resistance (Napolitano‐Tabare et al, 2021; Poddar et al, 2021; Tao et al, 2015). The large biomolecules like nucleic acids get sufficient surface area to bind with the column, and bound nucleic acids can easily get eluted out simply by changing column conditions like pH or ionic strength.…”

Protein Attachment Mechanism for Improved Functionalization of Affinity Monolith Chromatography (AMC)

“…In the present work, the affinity monolith chromatography (AMC) potential of previously fabricated and functionalized amine-modified alumina monolithic columns was analyzed [ 26 , 27 ]. A significant amount of literature is available about the surface properties and interaction behavior of proteins with monolith supports [ 28 , 29 , 30 ], but not so much about protein attachment mechanisms in immunoaffinity-based chromatography. In addition, since the interactions are mainly related to the type of support and surface properties, the kind of immobilization strategy, and the biomolecule properties (tendency to aggregate, etc.…”