Affinity chromatography supporting matrix

Sinninger, V. Tapon-Bretaudiere, J. Zhou, F.L. Bros, A. et al. Immobihzation of dermatan sulphate on a sihca matrix and its possible use as an affinity chromatography support for heparin cofactor II purification. J. Chromatogr. 1991, 539, 289. 

Metal-chelate affinity chromatography is a powerful purification technique whereby proteins or other molecules can be separated based upon their ability to form coordination complexes with immobilized metal ions (Porath et al., 1975 Lonnerdal and Keen, 1982 Porath and Belew, 1983 Porath and Olin, 1983 Sulkowski, 1985 Kagedal, 1989). The metal ions are stabilized on a matrix through the use of chelating compounds which usually have multivalent points of interaction with the metal atoms. To form useful affinity supports, these metal ion complexes must have some free or weakly associated and exchangeable coordination sites. These exchangeable sites then can form complexes with coordination sites on proteins or other molecules. Substances that are able to interact with the immobilized metals will bind and be retained on... 

Figure 6.14 Schematic representation of the principle of biospecific affinity chromatography. The chosen affinity ligand is chemically attached to the support matrix (agarose bead) via a suitable spacer arm. Only those ligands in solution that exhibit biospecific affinity for the immobilized species will be retained...

Selection of the matrix used to immobilize a ligand requires consideration of several properties. The stationary supports used in gel exclusion chromatography are found to be quite suitable for affinity chromatography because (1) they are physically and chemically stable under most experimental conditions, (2) they are relatively free of nonspecific adsorption effects, (3) they have satisfactory flow characteristics, (4) they are available with very large pore sizes, and (5) they have reactive functional groups to which an appropriate ligand may be attached. 

Affinity chromatography (AC) is a fractionation technique widely used in targeted proteomics or protein-protein interaction approaches. This method utilizes an interaction or affinity of a target protein to a substrate (or another protein) immobilized on a support matrix. Figure 17.4 demonstrates that AC can be incorporated into 3D proteomic approaches (Lee and Lee, 2004). Traditionally, AC has been used to purify, for example, carbohydratebinding proteins from Diplostomum pseudo-spathaceum (Mikes and Man, 2003) and the detoxification superfamily glutathione transferase (GST) from parasitic flatworms (Brophy and Barrett, 1990), specifically shown in... 

The basis for selectivity in affinity chromatography is the use of immobilized biochemicals, known as affinity ligands, that are covalently attached to a support matrix, as illustrated in Figure 2.17. The primary criteria that govern the suitability of a support matrix for affinity chromatography include (1) the mechanical and flow properties of the matrix, (2) the ease of covalent coupling of the ligand to the matrix, and (3) the stability of the... 

These biopolymers can be used for the immobilization of metal ions not only with the final objective of metal recovery (and subsequent valorization by desorption or chemical/thermal destmction of the polymer matrix) but also for elaborating new materials or designing new applications. Depending on the metal immobilized on the biopolymer, it is possible to design new sorbents (immobilization of iron on alginate [119], of molybdate on chitosan [59], for As(V) removal, of silver on chitosan for pesticide removal [120]), supports for affinity chromatography [121], antimicrobial material [122], drug release material [123], neutron capture therapy [124], and photoluminescent materials [125]. These are only a few... 

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