title: Profiling of serum antibodies with printed glycan array: room for data misinterpretation.
abstract: Using an example of Galβ1-3GlcNAc (Le(C)) related glycans, we here demonstrate a risk of data misinterpretation when polyclonal antibodies are probed for their glycan-binding specificities with help of a printed glycan array (PGA). Affinity isolation of antibodies from human serum using Le(C)-Sepharose or 3'-O-SuLe(C)-Sepharose in conditions of excess of the adsorbents generated identical material regardless of the affinity ligand, with the antibodies equally capable of binding to Le(C) and to 3'-O-SuLe(C) disaccharides, as well as to 3'-O-SiaLe(C) trisaccharide. More detailed profiling has shown that the isolated antibodies bind to the inner part of Galβ1-3GlcNAc disaccharide. We therefore conclude that serum does not contain different subsets of antibodies specific either to Le(C) or to 3'-O-SuLe(C), despite their visibly different binding signals to these glycans on PGA.
authors: Shemyakin-Ovchinnikov Institute of Bioorganic Chemistry, Russian Academy of Sciences, Miklukho-Maklaya 16/10, 117997 Moscow, Russia.
authors: Polina
authors: P
authors: Obukhova
authors: Vladimir
authors: V
authors: Piskarev
authors: Vyacheslav
authors: V
authors: Severov
authors: Galina
authors: G
authors: Pazynina
authors: Alexander
authors: A
authors: Tuzikov
authors: Maxim
authors: M
authors: Navakouski
authors: Nadezhda
authors: N
authors: Shilova
authors: Nicolai
authors: N
authors: Bovin
mesh_terms: D000117:Acetylglucosamine / Q000737:chemistry / Q000276:immunology; D000328:Adult; D000906:Antibodies / Q000097:blood* / Q000302:isolation & purification; D000915:Antibody Affinity / Q000276:immunology; D000918:Antibody Specificity / Q000276:immunology; D002240:Carbohydrate Sequence; D003627:Data Interpretation, Statistical; D004797:Enzyme-Linked Immunosorbent Assay; D000939:Epitopes / Q000276:immunology; D006801:Humans; D007983:Lewis Blood Group Antigens / Q000276:immunology; D046228:Microarray Analysis / Q000379:methods*; D008958:Models, Molecular; D008969:Molecular Sequence Data; D011134:Polysaccharides / Q000737:chemistry / Q000276:immunology*; D013431:Sulfates / Q000276:immunology; D014019:Tissue Donors
chemical_list: D000906:Antibodies; D000939:Epitopes; D007983:Lewis Blood Group Antigens; D011134:Polysaccharides; D013431:Sulfates; D000117:Acetylglucosamine