RRC ID 18649
著者 Takegawa Y, Araki K, Fujitani N, Furukawa J, Sugiyama H, Sakai H, Shinohara Y.
タイトル Simultaneous analysis of heparan sulfate, chondroitin/dermatan sulfates, and hyaluronan disaccharides by glycoblotting-assisted sample preparation followed by single-step zwitter-ionic-hydrophilic interaction chromatography.
ジャーナル Anal Chem
Abstract Glycosaminoglycans (GAGs) play important roles in cell adhesion and growth, maintenance of extracellular matrix (ECM) integrity, and signal transduction. To fully understand the biological functions of GAGs, there is a growing need for sensitive, rapid, and quantitative analysis of GAGs. The present work describes a novel analytical technique that enables high throughput cellular/tissue glycosaminoglycomics for all three families of uronic acid-containing GAGs, hyaluronan (HA), chondroitin sulfate (CS)/dermatan sulfate (DS), and heparan sulfate (HS). A one-pot purification and labeling procedure for GAG Δ-disaccharides was established by chemo-selective ligation of disaccharides onto high density hydrazide beads (glycoblotting) and subsequent labeling by fluorescence. The 17 most common disaccharides (eight comprising HS, eight CS/DS, and one comprising HA) could be separated with a single chromatography for the first time by employing a zwitter-ionic type of hydrophilic-interaction chromatography column. These novel analytical techniques were able to precisely characterize the glycosaminoglycome in various cell types including embryonal carcinoma cells and ocular epithelial tissues (cornea, conjunctiva, and limbus).
巻・号 83(24)
ページ 9443-9
公開日 2011-12-15
DOI 10.1021/ac2021079
PMID 22044073
MeSH Animals Cell Line Chondroitin Sulfates / analysis* Chromatography, High Pressure Liquid* Conjunctiva / metabolism Cricetinae Dermatan Sulfate / analysis* Disaccharides / analysis Heparitin Sulfate / analysis* Humans Hyaluronic Acid / analysis* Hydrophobic and Hydrophilic Interactions Ions / chemistry Mice Rabbits
IF 6.785
引用数 33
WOS 分野 CHEMISTRY, ANALYTICAL
リソース情報
ヒト・動物細胞 CHO-K1(RCB0285)