Affinity chromatography ligand selection

Affinity chromatography using factor XII as ligand leads to purification of u-PAR rather selectively, with only trace quantities of cytokeratin 1 or gClqR present [K. Joseph and A. Kaplan, unpubl. observations]. It is of interest that none of these three proteins possesses a transmembrane domain but u-PAR has a phos-phatidylinositol linkage within the cell membrane. Nevertheless, each of them has been isolated from purified cell membranes and they have been demonstrated to exist within the cell membrane by immunoelectron microscopy [41] presumably... 

Affinity chromatography exploits the high selectivity of most proteins for their ligands. Enzymes may be puri-... 

N.E. Katsos, N.E. Labrou and Y.D. Clonis, Interaction of L-glutamate oxidase with triazine dyes selection of ligands for affinity chromatography. J. Chromatogr.B, 807 (2004) 277-285. 

Table 3.19. Some forms of affinity chromatography that may be employed to purify a selected protein. Virtually all proteins carry out their biological effects by interacting in some way with other molecules, which can be given the general title ligands . This interaction is often quite biospecific. Immobilization of such ligands (or molecules which mimic ligands) onto chromatographic beads thus facilitates selective purification of the molecule of interest...

These adsorbants are based on immobilised ligands, which have a high affinity for a particular analyte (Fig. 15.13). There are examples where antibodies have been raised to an analyte and then bound to the surface of a SPE matrix. Various types of chemistry permit this type of immobilisation and affinity chromatography is well established in biochemistry. With the proliferation with biotechnological products such as therapeutic peptides, the use of these types of columns for extraction may increase since they can be designed to be highly selective for such compounds. 

Although its use in ligand selection for large scale of affinity chromatography is not wide, ribosome display and systematic evolution of ligands by exponential enrichment (SELEX) may... 

Labrou NE. Design and selection of ligands for affinity chromatography. Journal of Chromatography B 2003 790 67-78. 

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