title: Preparation of Glycan Arrays Using Pyridylaminated Glycans.
abstract: We describe the method to prepare neoglycoproteins from the conjugation of bovine serum albumin and pyridylaminated glycans. Large quantities of glycans (>1 mg) can be pyridylaminated and then converted to their 1-amino-1-deoxy derivatives by reaction with hydrogen followed by hydrazine. These pyridylaminated glycans can then be conjugated to bovine serum albumin via esterification with N-( m-maleimidobenzoyloxy)succinimide to form a neoglycoprotein, e.g., glycosylated bovine serum albumin. As a demonstration, we prepared High-mannose bovine serum albumin, which was immobilized on an activated glass slide. Then, we showed that the neoglycoprotein bind to Cy3-labeled Lens culinaris agglutinin, a mannose-specific plant lectin, as detected using an evanescent-field-activated fluorescence scanner system.
authors.affiliation: Department of Functional Glycomics, Life Science Research Center, Kagawa University, 1750-1 Ikenobe, Miki-cho, Kita-gun, Kagawa, 761-0793, Japan. nakakita@med.kagawa-u.ac.jp.
authors.forename: Shin-Ichi
authors.initials: S
authors.lastname: Nakakita
authors.affiliation: Department of Functional Glycomics, Life Science Research Center, Kagawa University, 1750-1 Ikenobe, Miki-cho, Kita-gun, Kagawa, 761-0793, Japan.
authors.forename: Jun
authors.initials: J
authors.lastname: Hirabayashi
mesh_terms: D000586:Amination; D000818:Animals; D002417:Cattle; D006023:Glycoproteins / Q000737:chemistry / Q000378:metabolism; D046228:Microarray Analysis / Q000379:methods*; D011134:Polysaccharides / Q000737:chemistry / Q000378:metabolism*; D011725:Pyridines / Q000737:chemistry*; D012710:Serum Albumin, Bovine / Q000378:metabolism
chemical_list: D006023:Glycoproteins; D011134:Polysaccharides; D011725:Pyridines; D012710:Serum Albumin, Bovine; C023666:pyridine
keywords: 1-Amino-1-deoxy derivative; Evanescent-field-activated fluorescence scanner; Glycan array; Neoglycoprotein; Pyridylamination