title: Versatile glycoblotting nanoparticles for high-throughput protein glycomics.
abstract: We have developed an effective and practical trap-and-release method based on chemoselective ligation of carbohydrates with reactive aminooxyl groups attached to the surface of nanoparticles (referred to as glycoblotting nanoparticles). These glycoblotting nanoparticles were synthesized by UV irradiation of diacetylene-functionalized lipids that contain the aminooxyl group. The glycoblotting nanoparticles captured carbohydrates in aqueous solution under mild conditions and were collected by simple centrifugation. The trapped carbohydrates were effectively released from the nanoparticles under acidic conditions to give pure oligosaccharides. This glycoblotting process reduced the time required for the purification process of carbohydrates to less than 6 h, compared to the several days needed for conventional chromatographic techniques. The oligosaccharides (N-glycan) were released from ovalbumin (glycoprotein) by PNGase F after tryptic digestion. MALDI-TOF mass spectra before purification did not show any significant signals corresponding to N-glycans because these signals were hidden by the large signals of the abundant peptides. However, after purification with the glycoblotting nanoparticles, only signals corresponding to oligosaccharides appeared. We also demonstrated a clear analysis of the oligosaccharides contained in the mice dermis by means of glycoblotting.
authors.affiliation: Division of Biological Sciences, Graduate School of Science, Frontier Research Center for Post-Genome Science and Technology, Hokkaido University, Kita , Kita-ku, Sapporo 001-0021, Japan.
authors.forename: Kenichi
authors.initials: K
authors.lastname: Niikura
authors.forename: Ryosuke
authors.initials: R
authors.lastname: Kamitani
authors.forename: Masaki
authors.initials: M
authors.lastname: Kurogochi
authors.forename: Rie
authors.initials: R
authors.lastname: Uematsu
authors.forename: Yasuro
authors.initials: Y
authors.lastname: Shinohara
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Nakagawa
authors.forename: Kisaburo
authors.initials: K
authors.lastname: Deguchi
authors.forename: Kenji
authors.initials: K
authors.lastname: Monde
authors.forename: Hirosato
authors.initials: H
authors.lastname: Kondo
authors.forename: Shin-Ichiro
authors.initials: S
authors.lastname: Nishimura
mesh_terms: D000818:Animals; D002845:Chromatography; D006020:Glycopeptides / Q000032:analysis* / Q000737:chemistry; D008081:Liposomes / Q000737:chemistry; D051379:Mice; D049329:Nanostructures / Q000737:chemistry*; D009844:Oligosaccharides / Q000032:analysis / Q000737:chemistry; D010777:Photochemistry; D012867:Skin / Q000737:chemistry; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D013997:Time Factors
chemical_list: D006020:Glycopeptides; D008081:Liposomes; D009844:Oligosaccharides