title: Neoglycolipid-based "designer" oligosaccharide microarrays to define β-glucan ligands for Dectin-1.
abstract: In this chapter, we describe the key steps of the "designer" oligosaccharide microarray approach we followed to prove the carbohydrate binding activity and define the oligosaccharide ligands for Dectin-1, an atypical C-type lectin-like signaling receptor of the mammalian innate immune system with a key role in anti-fungal immunity. The term "designer" microarray, which we introduced in the course of the Dectin-1 study refers to a microarray of oligosaccharide probes generated from ligand-bearing glycoconjugates to reveal the oligosaccharide ligands they harbor, so that these can be isolated and characterized. Oligosaccharide probes were generated from two polysaccharides, one that was bound by Dectin-1 and known to be rich in β1,3-glucose sequence and another that was not bound and was rich in β1,6-glucose sequence and served as a negative control. The approach involved: classic ELISA-type binding assays to select the polysaccharides; partial depolymerization of the polysaccharides by chemical hydrolysis; fractionation by size of the glucan oligosaccharides obtained and determination of their chain lengths by mass spectrometry; detection of Dectin-1 ligand-positive and ligand-negative oligosaccharides using the neoglycolipid (NGL) technology; methylation analysis of oligosaccharides to derive glucose linkage information, and incorporation of the newly generated glucan oligosaccharide probes into microarrays encompassing diverse mammalian-type and exogenous sequences for microarray analysis of Dectin-1.
authors: The Glycosciences Laboratory, Department of Medicine, Imperial College London, Hammersmith Campus, London, UK. maria.palma@dg.fct.unl.pt
authors: Angelina S
authors: AS
authors: Palma
authors: Yibing
authors: Y
authors: Zhang
authors: Robert A
authors: RA
authors: Childs
authors: Maria A
authors: MA
authors: Campanero-Rhodes
authors: Yan
authors: Y
authors: Liu
authors: Ten
authors: T
authors: Feizi
authors: Wengang
authors: W
authors: Chai
mesh_terms: D005936:Glucans / Q000737:chemistry*; D006017:Glycolipids / Q000737:chemistry*; D037181:Lectins, C-Type / Q000737:chemistry*; D008024:Ligands; D046228:Microarray Analysis / Q000379:methods*; D000099181:Dectin-1
chemical_list: D005936:Glucans; D006017:Glycolipids; D037181:Lectins, C-Type; D008024:Ligands; D000099181:Dectin-1