title: A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates.
abstract: Because sugar and its derivatives play important roles in various biological phenomena, the rapid and high-throughput analysis of various glycoconjugates is keenly desirable. We describe herein the construction of a novel fluorescent lectin array for saccharide detection using a supramolecular hydrogel matrix. In this array, the fluorescent lectins were noncovalently fixed under semi-wet conditions to suppress the protein denaturation. It is demonstrated by fluorescence titration and fluorescence lifetime experiments that the immobilized lectins act as a molecular recognition scaffold in the hydrogel matrix, similar to that in aqueous solution. That is, a bimolecular fluorescence quenching and recovery (BFQR) method can successfully operate under both conditions. This enables one to fluorescently read-out a series of saccharides on the basis of the recognition selectivity and affinity of the immobilized lectins without tedious washing processes and without labeling the target saccharides. Simple and high-throughput sensing and profiling were carried out using the present lectin array for diverse glycoconjugates, which not only included a simple glucose, but also oligosaccharides, and glycoproteins, and, furthermore, the pattern recognition and profiling of several types of cell lysates were also accomplished.
authors.affiliation: PRESTO (Synthesis and Control, JST), Department of Synthetic Chemistry and Biological Chemistry, Kyoto University, Kyotodaigaku-Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
authors.forename: Yoichiro
authors.initials: Y
authors.lastname: Koshi
authors.forename: Eiji
authors.initials: E
authors.lastname: Nakata
authors.forename: Hiroki
authors.initials: H
authors.lastname: Yamane
authors.forename: Itaru
authors.initials: I
authors.lastname: Hamachi
mesh_terms: D005456:Fluorescent Dyes / Q000737:chemistry*; D006001:Glycoconjugates / Q000032:analysis*; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate / Q000737:chemistry*; D037102:Lectins / Q000737:chemistry*; D015394:Molecular Structure; D040081:Protein Array Analysis
chemical_list: D005456:Fluorescent Dyes; D006001:Glycoconjugates; D037102:Lectins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate