title: Using Single Lectins to Enrich Glycoproteins in Conditioned Media.
abstract: Lectins are sugar-binding proteins that can recognize and bind to carbohydrates conjugated to proteins and lipids. Coupled with mass spectrometry technologies, lectin affinity chromatography is becoming a popular approach for identification and quantification of glycoproteins in complex samples such as blood, tumor tissues, and cell lines. Given the commercial availability of a large number of lectins that recognize diverse sugar structures, it is now possible to isolate and study glycoproteins for biological and medical research. This unit provides a general guide to single-lectin-based enrichment of glycoproteins from serum-free conditioned media. Due to the unique carbohydrate specificity of most lectins and the complexity of the samples, optimization steps may be required to evaluate different elution buffers and methods as well as binding conditions, for each lectin, for optimal recovery of bound glycoproteins.
authors.affiliation: Department of Chemistry and Biomolecular Sciences, Macquarie University, Sydney, Australia.
authors.forename: Manveen K
authors.initials: MK
authors.lastname: Sethi
authors.affiliation: Australian School of Advanced Medicine, Macquarie University, Sydney, Australia.
authors.forename: Susan
authors.initials: S
authors.lastname: Fanayan
mesh_terms: D002460:Cell Line; D002846:Chromatography, Affinity / Q000379:methods*; D017077:Culture Media, Conditioned / Q000737:chemistry*; D006023:Glycoproteins / Q000032:analysis / Q000737:chemistry / Q000302:isolation & purification* / Q000378:metabolism*; D006801:Humans; D037102:Lectins / Q000737:chemistry / Q000378:metabolism*; D053719:Tandem Mass Spectrometry
chemical_list: D017077:Culture Media, Conditioned; D006023:Glycoproteins; D037102:Lectins
keywords: cell line; conditioned media; glycoprotein enrichment; lectin