Fractionation of membrane proteins on immobilized lectins by high-performance liquid affinity chromatography.
Detergent-solubilized glycoproteins were fractionated on high-performance affinity columns employing A concanavalin and Pisum sativum agglutinin as ligands immobilized on microparticulate silica via a propyl spacer. The separations were characterized by high recovery (90-95%) and high specificity (enrichment factor 6- and 58-fold for the bound fractions on A concanavalin and P. sativum agglutinin columns, respectively, compared with the crude extract), as estimated by enzyme-linked lectin assay and chromatographic criteria. In addition, the short running times (30-40 min) make this method highly useful for a first characterization of complex glycoprotein samples and of individual molecules.
D002846:Chromatography, Affinity / Q000379:methods; D002851:Chromatography, High Pressure Liquid / Q000379:methods; D003208:Concanavalin A; D005260:Female; D005347:Fibroblasts / Q000032:analysis; D006023:Glycoproteins / Q000032:analysis; D006801:Humans; D037102:Lectins; D008168:Lung / Q000196:embryology; D008565:Membrane Proteins / Q000032:analysis*; D037121:Plant Lectins; D011247:Pregnancy
D006023:Glycoproteins; D037102:Lectins; D008565:Membrane Proteins; D037121:Plant Lectins; C036292:pea lectin; D003208:Concanavalin A