Chromatography of lectins

Hydrophilic water-insoluble gels suitable for affinity chromatography of lectins have been prepared by co-polymerization of acrylamide, A/, AT-methyIene bisacrylamide and alkenyl l-thioglycosides. ... 

Lectin affinity chromatography may be used to purify a range of glycoproteins. Lectins are a group of proteins synthesized by plants, vertebrates and a number of invertebrate species. Especially high levels of lectins are produced by a variety of plant seeds. Plant lectins are often termed... 

Figure 2.18 Wheat germ lectin isolation using A -acetyl-o-glucosamine as the ligand. Chromatography conditions column, Waters API glass column, 100 x 10 mm I.D. flow rate, 1 ml/min detection, UV absorbance at 280 nm. The sample was 1 ml of crude wheat germ preparation 1.2 mg/ml of lectin was eluted from a crude preparation with 6.8 mg/ml total protein. (Reprinted from Ref. 64 with permission.)...

A mixture of lentil lectin-reactive glycoproteins from pig lymphocyte-plasma membrane was isolated by lentil lectin-Sepharose chromatography of sodium deoxycholate-solubilized membranes.445 Eighty-seven percent of the protein applied (17% of hexose) passed through unretarded, and 13% of the applied protein (83% of hexose) was bound, and eluted with methyl a-D-glucopyranoside solution. Recovery was 95% of the material applied, in contrast to the recovery in similar experiments conducted on con A-Sepharose columns (80% recovery).850 The eluate from the lentil column, which contained at least ten glycoproteins, blocked lymphocyte transformation induced by lentil or kidney-bean lectins.445... 

Nearly all current methods of isolation and purification of lectins rely on affinity chromatography. Naturally, the characteristic ligand must be determined in advance. The properties of lectins can be used to precipitate macromolecules and to agglutinate some types of cells, be they plant or animal. The driving force of this reaction is the association with certain bound residues, generally monosaccharides, from the macromolecule or the cellular periphery. When this type of reaction is observed, the problem is to find the sugar that can inhibit activity at the lowest possible molar concentration. As in the case of immunochemical precipitations, this inhibition is due to the occupation of the recognition site by the small soluble molecule. 

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