Versatile strategy for the synthesis of biotin-labelled glycans, their immobilization to establish a bioactive surface and interaction studies with a lectin on a biochip

• Published in: Glycoconjugate journal Vol. 25; no. 7; pp. 633 - 646
• Main Authors: Muñoz, F. Javier, Rumbero, Ángel, Sinisterra, José V, Santos, J. Ignacio, André, Sabine, Gabius, Hans-J, Jiménez-Barbero, Jesús, Hernáiz, María J
• Format: Journal Article
• Language: English
• Published: Boston Boston : Springer US 01-10-2008; Springer US; Springer Nature B.V
• Subjects: Amination; Biochemistry; Biomedical and Life Sciences; Biosynthesis; Biotin - chemistry; Biotinylation; Carbohydrate Conformation; Carbohydrate-lectin interactions; Carbohydrates; Cellular biology; Fluorescent Dyes; Glycan labelling; Glycoconjugates - chemical synthesis; Glycoconjugates - chemistry; Immobilization of glycans; Kinetics; Life Sciences; Magnetic Resonance Spectroscopy; Mistletoe - chemistry; Models, Molecular; Molecular recognition; NMR; Nuclear magnetic resonance; Pathology; Plant Lectins - chemistry; Plant Lectins - metabolism; Polysaccharides - chemical synthesis; Polysaccharides - chemistry; Protein Array Analysis; SPR; Surface Plasmon Resonance - methods; Molecular recognition; NMR; SPR; Carbohydrate-lectin interactions; Immobilization of glycans; Glycan labelling
• ISSN: 0282-0080; 1573-4986
• DOI: 10.1007/s10719-008-9115-y

The emerging role of glycans as versatile biochemical signals in diverse aspects of cellular sociology calls for establishment of sensitive methods to monitor carbohydrate recognition by receptors such as lectins. Most of these techniques involve the immobilization of one of the binding partners on a surface, e.g. atomic force microscopy, glycan array and Surface Plasmon Resonance (SPR), hereby simulating cell surface presentation. Here, we report the synthesis of fluorescent glycoconjugates, with a functionalization strategy which avoids the frequently occurring ring opening at the reducing end for further immobilization on a surface or derivatization with biotin. In order to improve the versatility of these derivatized glycans for biological studies, a new approach for the synthesis of biotinylated and fluorescent glycans has also been realized. Finally, to illustrate their usefulness the neoglycoconjugates were immobilized on different surfaces, and the interaction analysis with a model lectin, the toxin from mistletoe, proved them to act as potent ligands, underscoring the merit of the presented synthetic approach.