title: Fabrication of Carbohydrate Microarrays by Boronate Formation.
abstract: The interactions between soluble carbohydrates and/or surface displayed glycans and protein receptors are essential to many biological processes and cellular recognition events. Carbohydrate microarrays provide opportunities for high-throughput quantitative analysis of carbohydrate-protein interactions. Over the past decade, various techniques have been implemented for immobilizing glycans on solid surfaces in a microarray format. Herein, we describe a detailed protocol for fabricating carbohydrate microarrays that capitalizes on the intrinsic reactivity of boronic acid toward carbohydrates to form stable boronate diesters. A large variety of unprotected carbohydrates ranging in structure from simple disaccharides and trisaccharides to considerably more complex human milk and blood group (oligo)saccharides have been covalently immobilized in a single step on glass slides, which were derivatized with high-affinity boronic acid ligands. The immobilized ligands in these microarrays maintain the receptor-binding activities including those of lectins and antibodies according to the structures of their pendant carbohydrates for rapid analysis of a number of carbohydrate-recognition events within 30 h. This method facilitates the direct construction of otherwise difficult to obtain carbohydrate microarrays from underivatized glycans.
authors.affiliation: Department of Chemistry, National Tsing Hua University, 101, Sec 2 Kuang-Fu Rd, Hsinchu, 30013, Taiwan. aka_oc@yahoo.co.in.
authors.forename: Avijit K
authors.initials: AK
authors.lastname: Adak
authors.affiliation: Department of Chemistry, National Tsing Hua University, 101, Sec 2 Kuang-Fu Rd, Hsinchu, 30013, Taiwan.
authors.forename: Ting-Wei
authors.initials: TW
authors.lastname: Lin
authors.affiliation: Department of Chemistry, National Tsing Hua University, 101, Sec 2 Kuang-Fu Rd, Hsinchu, 30013, Taiwan.
authors.forename: Ben-Yuan
authors.initials: BY
authors.lastname: Li
authors.affiliation: Department of Chemistry, National Tsing Hua University, 101, Sec 2 Kuang-Fu Rd, Hsinchu, 30013, Taiwan. cclin66@mx.nthu.edu.tw.
authors.forename: Chun-Cheng
authors.initials: CC
authors.lastname: Lin
mesh_terms: D001897:Boronic Acids / Q000737:chemistry*; D002241:Carbohydrates / Q000737:chemistry*; D046228:Microarray Analysis / Q000379:methods*; D011506:Proteins / Q000737:chemistry
chemical_list: D001897:Boronic Acids; D002241:Carbohydrates; D011506:Proteins
keywords: Boronate formation; Boronic acid; Carbohydrates; Carbohydrate–protein interactions; Glass slides; Glycan immobilization; Lectins; Microarrays