Fractionation of membrane proteins on immobilized lectins by high-performance liquid affinity chromatography

• Published in: Analytical biochemistry Vol. 151; no. 2; p. 424
• Main Authors: Renauer, D, Oesch, F, Kinkel, J, Unger, K K, Wieser, R J
• Format: Journal Article
• Language: English
• Published: United States 01-12-1985
• Subjects: Chromatography, Affinity - methods; Chromatography, High Pressure Liquid - methods; Concanavalin A; Female; Fibroblasts - analysis; Glycoproteins - analysis; Humans; Lectins; Lung - embryology; Membrane Proteins - analysis; Plant Lectins; Pregnancy
• ISSN: 0003-2697
• DOI: 10.1016/0003-2697(85)90198-8

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.