title: Examination of whole cell galectin binding by solid phase and flow cytometric analysis.
abstract: We have utilized simple flow cytometric and fluorescence-based solid phase assays to study the interaction of glycan-binding proteins (GBP) to cell surface glycoconjugates. These methods utilize commonly employed flow cytometry techniques and commercially available streptavidin-coated microplates to immobilize various biotinylated ligands, such as glycopeptides, oligosaccharides, and whole cells. Using this approach, fluorescently labeled GBPs, in particular, members of the galectin family, can be interrogated for potential interactions with cell surface carbohydrates, including elucidation of the potential impact of alterations in glycosylation on carbohydrate recognition. Using these approaches, we present examples of flow cytometric and fluorescence-based solid phase assays to study galectin-carbohydrate interactions.
authors.affiliation: Glykos Finland Ltd., Viikinkaari 6, 00790, Helsinki, Finland, Anne.Leppanen@helsinki.fi.
authors.forename: Anne
authors.initials: A
authors.lastname: Leppänen
authors.forename: Connie M
authors.initials: CM
authors.lastname: Arthur
authors.forename: Sean R
authors.initials: SR
authors.lastname: Stowell
authors.forename: Richard D
authors.initials: RD
authors.lastname: Cummings
mesh_terms: D000588:Amines / Q000737:chemistry; D019921:Biotinylation; D018914:Cells, Immobilized / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods*; D005456:Fluorescent Dyes / Q000737:chemistry*; D037161:Galectins / Q000737:chemistry* / Q000378:metabolism*; D006026:Glycoside Hydrolases / Q000378:metabolism; D018922:HL-60 Cells; D006801:Humans; D018551:Solanum lycopersicum / Q000737:chemistry; D037121:Plant Lectins / Q000737:chemistry / Q000378:metabolism; D011485:Protein Binding
chemical_list: D000588:Amines; D005456:Fluorescent Dyes; D037161:Galectins; C045475:Griffonia simplicifolia lectins; D037121:Plant Lectins; D006026:Glycoside Hydrolases