Immobilization and Clustering of Structurally Defined Oligosaccharides for Sugar Chips:  An Improved Method for Surface Plasmon Resonance Analysis of Protein−Carbohydrate Interactions

Abstract

Oligosaccharides are increasingly being recognized as important partners in receptor-ligand binding and cellular signaling. Surface plasmon resonance (SPR) is a very powerful tool for the real-time study of the specific interactions between biological molecules. We report here an advanced method for the immobilization of oligosaccharides in clustered structures for SPR and their application to the analysis of heparin-protein interactions. Reductive amination reactions and linker molecules were designed and optimized. Using mono-, tri-, or tetravalent linker compounds, we incorporated synthetic structurally defined disaccharide units of heparin and immobilized them as ligands for SPR. Their binding to an important hemostatic protein, von Willebrand factor (vWf), and its known heparin-binding domain was quantitatively analyzed. These multivalent ligand conjugates exhibited reproducible binding behavior, with consistency of the surface conditions of the SPR chip. This novel technique for oligosaccharide immobilization in SPR studies is accurate, specific, and easily applicable to both synthetic and naturally derived oligosaccharides.

Staphylococcus aureus SasA Is Responsible for Binding to the Salivary Agglutinin gp340, Derived from Human Saliva

“…8; note that the saccharide unit of all sugar chains at the reducing end, which loses pyranose structure during the reductive amination reaction, serves as a hydrophilic spacer). To immobilize the sugar chains, the ligand conjugates were first prepared by reacting sugar chains with linker compounds and were immobilized on gold-coated chips, as previously reported (33). Briefly, gold-coated chips (Toyobo, Shiga, Japan) were washed with UV ozone cleaner (Structure Probe Inc., West Chester, PA).…”

Targeting the carbohydrates on HIV-1: Interaction of oligomannose dendrons with human monoclonal antibody 2G12 and DC-SIGN

“…Over the past several years, many groups have found that glycodendrimers/ glycodendrons exhibit strikingly enhanced affinity against target proteins (23,31,32). More recently, different synthetic approaches and applications of these macromolecules have been explored (33)(34)(35)(36).…”

Interaction Analyses of Amyloid β Peptide (1–40) with Glycosaminoglycan Model Polymers

“…The chemical structures of the GAGs are too complex to allow the details of their interaction with A¢ to be elucidated directly, and an ingenious method to analyze the interaction is needed. 7,8 We have reported use of sulfonated glycopolymers as mimic polymers of GAGs, to investigate the GAGsA¢ interaction. 9 The advantage of glycopolymers is facile preparation, so that the effects of saccharide structure and polymer molecular weight can be easily investigated.…”