title: Evanescent-field fluorescence-assisted lectin microarray: a new strategy for glycan profiling.
abstract: Glycans have important roles in living organisms with their structural diversity. Thus, glycomics, especially aspects involving the assignment of functional glycans in a high-throughput manner, has been an emerging field in the postproteomics era. To date, however, there has been no versatile method for glycan profiling. Here we describe a new microarray procedure based on an evanescent-field fluorescence-detection principle, which allows sensitive, real-time observation of multiple lectin-carbohydrate interactions under equilibrium conditions. The method allows quantitative detection of even weak lectin-carbohydrate interactions (dissociation constant, K(d) > 10(-6) M) as fluorescent signals for 39 immobilized lectins. We derived fully specific signal patterns for various Cy3-labeled glycoproteins, glycopeptides and tetramethylrhodamine (TMR)-labeled oligosaccharides. The obtained results were consistent with the previous reports of glycoprotein and lectin specificities. We investigated the latter aspects in detail by frontal affinity chromatography, another profiling method. Thus, the developed lectin microarray should contribute to creation of a new paradigm for glycomics.
authors.affiliation: Research Center for Glycoscience (RCG), National Institute of Advanced Industrial Science and Technology (AIST), AIST Tsukuba Central 2; Tsukuba, Ibaraki 305-8568, Japan.
authors.forename: Atsushi
authors.initials: A
authors.lastname: Kuno
authors.forename: Noboru
authors.initials: N
authors.lastname: Uchiyama
authors.forename: Shiori
authors.initials: S
authors.lastname: Koseki-Kuno
authors.forename: Youji
authors.initials: Y
authors.lastname: Ebe
authors.forename: Seigo
authors.initials: S
authors.lastname: Takashima
authors.forename: Masao
authors.initials: M
authors.lastname: Yamada
authors.forename: Jun
authors.initials: J
authors.lastname: Hirabayashi
mesh_terms: D000818:Animals; D005453:Fluorescence; D037102:Lectins / Q000737:chemistry*; D051379:Mice; D046228:Microarray Analysis / Q000379:methods* / Q000592:standards; D011134:Polysaccharides / Q000032:analysis* / Q000737:chemistry*; D015203:Reproducibility of Results
chemical_list: D037102:Lectins; D011134:Polysaccharides