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
| Autor: | Hiroaki Sugiyama, Jun-ichi Furukawa, Kayo Araki, Yasuhiro Takegawa, Hideaki Sakai, Yasuro Shinohara, Naoki Fujitani |
|---|---|
| Rok vydání: | 2011 |
| Předmět: |
Dermatan Sulfate
Disaccharides Dermatan sulfate Cell Line Analytical Chemistry Glycosaminoglycan Extracellular matrix Mice chemistry.chemical_compound Cricetinae Animals Humans Chondroitin Chondroitin sulfate Hyaluronic Acid Chromatography High Pressure Liquid Ions Chromatography Hydrophilic interaction chromatography Chondroitin Sulfates Heparan sulfate chemistry Biochemistry Heparitin Sulfate Rabbits Chromatography column Conjunctiva Hydrophobic and Hydrophilic Interactions |
| Zdroj: | Analytical Chemistry. 83:9443-9449 |
| ISSN: | 1520-6882 0003-2700 |
| Popis: | 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). |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|