Sepharose-lectin column chromatography

Sepharose-lectin column chromatography To purify glycoproteins or gly-copeptides that bind the particular lectin used. 

The P. vulgaris lectins have now been isolated by affinity chromatography on columns of Sepharose conjugated with porcine thyroglobu-lin 149,154 an(j fetuin,156 both of which glycoproteins bind the lectin. 

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... 

The glycosylation can further be demonstrated by affinity chromatography on a galactose-specific immobilized lectin. The test was done on a PNA-Sepharose column with native and modified chymotrypsins. The chymotrypsin-galactose peak was shifted compared to the native enzyme. All the enzyme molecules were modified it was possible to follow the modification of the physico-chemical properties of the derivatized enzyme without further purification. 

---