Affinity monolith chromatography

Abstract

The combined use of monolithic supports with selective affinity ligands as stationary phases has recently given rise to a new method known as affinity monolith chromatography (AMC). This review will discuss the basic principles behind AMC and examine the types of supports and ligands that have been employed in this method. Approaches for placing affinity ligands in monoliths will be considered, including methods based on covalent immobilization, biospecific adsorption, entrapment, and the formation of coordination complexes. Several reported applications will then be presented, such as the use of AMC for bioaffinity chromatography, immunoaffinity chromatography, immobilized metal-ion affinity chromatography, dye-ligand affinity chromatography, and biomimetic chromatography. Other applications that will be discussed are chiral separations and studies of biological interactions based on AMC.

Chromatographic Immunoassays: Strategies and Recent Developments in The Analysis of Drugs and Biological Agents

“…For example, the use of nanomaterials, such as gold or silver nanoparticles, and quantum dots should continue to lead to new or enhanced detection methods and formats for these methods [19,[85][86][87][88][93][94][95][96][97][98][99][100]109]. The utilization of alternative supports, such as affinity monoliths, is also expected to continue [110][111][112][113][114][115][116]. In addition, further research is anticipated in the development of miniaturized chromatographic immunoassays that can be employed in microfluidic devices and portable or disposable devices [19,[26][27][28]30,[82][83][84][85][86][87][88]100,113,116].…”

The Value of Fungal Protease Inhibitors in Affinity Chromatography

“…It is exceptional in purification science as a tool for highly selective isolation of antigens (drugs, hormones, peptides, proteins, viruses, cell components), antibodies, enzymes, sugars, glycoproteins and glycolipids, immunoglobulins, nucleotide-binding and metalbinding peptides and proteins (Mallik & Hage, 2006). The use of specific interactions makes affinity chromatography a powerful tool in isolation of a particular substance from a complex mixture.…”

“…The ligand can consist of an immobilized sequence of DNA or RNA, a protein or enzyme, lectin, amino acids, immunoglobulin, a biomimetic dye, an enzyme substrate or inhibitor, or a small molecule (Hage, 2006;Healthcare, 2007). Target molecules to be purified are applied in a mobile phase known as the application buffer to the column containing insoluble polymer or gel.…”

“…Target molecules to be purified are applied in a mobile phase known as the application buffer to the column containing insoluble polymer or gel. The buffer is generally chosen to allow optimal binding of the immobilized ligand to its target while other sample components pass through with little or no retention (Mallik & Hage, 2006). Interaction between the ligand and the target molecule in a sample can be the result of electrostatic or hydrophobic interactions, van der Waals' forces or hydrogen bonding.…”

Evaluation of photografted charged sites within polymer monoliths in capillary columns using contactless conductivity detection

“…Examples include the immobilisation of protein A, for the separation of immunoglobulins [6][7][8], lectins such as concavalin A, for the extraction of glycoproteins [9], enzymes such as trypsin, for use as microbioreactors in LC-MS proteomic applications [4], and the immobilisation of immunoglobulins [10]. These varied applications of polymeric monolithic stationary phases for affinity chromatography has been the subject of a recent review by Mallik [11] in which the numerous immobilisation strategies which have previously been reported are discussed. The most common approaches result in covalent attachment of the protein to a monolith and include the epoxy method [12,13], the Schiff base method [7,10,12], the glutaraldehyde method [7,14], the carbonyldiimidazole method [7,10,12], the disuccinimidyl method, the hydrazide method [10], and the cyanogen bromide method.…”