title: Epitope detection chromatography: a method to dissect the structural heterogeneity and inter-connections of plant cell-wall matrix glycans.
abstract: Plant cell walls are complex, multi-macromolecular assemblies of glycans and other molecules and their compositions and molecular architectures vary extensively. Even though the chemistry of cell-wall glycans is now well understood, it remains a challenge to understand the diversity of glycan configurations and interactions in muro, and how these relate to changes in the biological and mechanical properties of cell walls. Here we describe in detail a method called epitope detection chromatography analysis of cell-wall matrix glycan sub-populations and inter-connections. The method combines chromatographic separations with use of glycan-directed monoclonal antibodies as detection tools. The high discrimination capacity and high sensitivity for the detection of glycan structural features (epitopes) provided by use of established monoclonal antibodies allows the study of oligosaccharide motifs on sets of cell-wall glycans in small amounts of plant materials such as a single organ of Arabidopsis thaliana without the need for extensive purification procedures. We describe the use of epitope detection chromatography to assess the heterogeneity of xyloglucan and pectic rhamnogalacturonan I sub-populations and their modulation in A. thaliana organs.
authors.affiliation: Faculty of Biological Sciences, Centre for Plant Sciences, University of Leeds, Leeds, LS2 9JT, UK.
authors.forename: Valérie
authors.initials: V
authors.lastname: Cornuault
authors.forename: Iain W
authors.initials: IW
authors.lastname: Manfield
authors.forename: Marie-Christine
authors.initials: MC
authors.lastname: Ralet
authors.forename: J Paul
authors.initials: JP
authors.lastname: Knox
mesh_terms: D000911:Antibodies, Monoclonal / Q000276:immunology; D017360:Arabidopsis / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D029681:Arabidopsis Proteins / Q000235:genetics / Q000378:metabolism; D002473:Cell Wall / Q000737:chemistry*; D002845:Chromatography / Q000379:methods*; D000939:Epitopes / Q000032:analysis* / Q000276:immunology; D005936:Glucans / Q000032:analysis / Q000737:chemistry; D010368:Pectins / Q000032:analysis / Q000737:chemistry; D010430:Pentosyltransferases / Q000235:genetics / Q000378:metabolism; D018517:Plant Roots / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D018520:Plant Shoots / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D010944:Plants / Q000737:chemistry*; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry / Q000276:immunology; D015203:Reproducibility of Results; D014990:Xylans / Q000032:analysis / Q000737:chemistry
chemical_list: D000911:Antibodies, Monoclonal; D029681:Arabidopsis Proteins; D000939:Epitopes; D005936:Glucans; D011134:Polysaccharides; D014990:Xylans; C042491:rhamnogalacturonan I; C029353:xyloglucan; D010368:Pectins; C509629:ARAD1 protein, Arabidopsis; C576737:ARAD2 protein, Arabidopsis; D010430:Pentosyltransferases
keywords: Arabidopsis thaliana; matrix glycans; pectin; plant cell walls; rhamnogalacturonan I; technical advance; xyloglucan