title: Three-dimensional arrays using glycoPEG tags: glycan synthesis, purification and immobilisation.
abstract: Glycan arrays have become the premier tool for rapidly establishing the binding or substrate specificities of lectins and carbohydrate-processing enzymes. New approaches for accelerating carbohydrate synthesis to address the enormous complexity of natural glycan structures are necessary. Moreover, optimising glycan immobilisation is key for the development of selective, sensitive and reproducible array-based assays. We present a tag-based approach that accelerates the preparation of glycan arrays on all levels by improving the synthesis, the purification and immobilisation of oligosaccharides. Glycan primers were chemically attached to bifunctional polyethyleneglycol (PEG) tags, extended enzymatically with the help of recombinant glycosyltransferases and finally purified by ultrafiltration. When printed directly onto activated glass slides, these glycoPEG tags afforded arrays with exceptionally high sensitivity, low background and excellent spot morphology. Likewise, the conjugation of glycoPEG tags to latex nanoparticles yielded multivalent scaffolds for carbohydrate-binding assays with very low non-specific binding.
authors: Biofunctional Nanomaterials Department, CICbiomaGUNE, Paseo Miramon 182, 20009 San Sebastian, Spain.
authors: Juan
authors: J
authors: Etxebarria
authors: Sonia
authors: S
authors: Serna
authors: Ana
authors: A
authors: Beloqui
authors: Manuel
authors: M
authors: Martin-Lomas
authors: Niels-Christian
authors: NC
authors: Reichardt
mesh_terms: D016695:Glycosyltransferases / Q000235:genetics / Q000378:metabolism*; D037102:Lectins / Q000737:chemistry*; D011092:Polyethylene Glycols / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism; D011134:Polysaccharides / Q000138:chemical synthesis* / Q000737:chemistry / Q000378:metabolism
chemical_list: D037102:Lectins; D011134:Polysaccharides; D011092:Polyethylene Glycols; D016695:Glycosyltransferases