A fluorescent lectin array using supramolecular hydrogel for simple detection and pattern profiling for various glycoconjugates.
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.
Koshi
Yoichiro
Y
PRESTO (Synthesis and Control, JST), Department of Synthetic Chemistry and Biological Chemistry, Kyoto University, Kyotodaigaku-Katsura, Nishikyo-ku, Kyoto, 615-8510, Japan.
Nakata
Eiji
E
Yamane
Hiroki
H
Hamachi
Itaru
I
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
D005456:Fluorescent Dyes; D006001:Glycoconjugates; D037102:Lectins; D020136:Hydrogel, Polyethylene Glycol Dimethacrylate