Analysis of glycoprotein-derived oligosaccharides in glycoproteins detected on two-dimensional gel by capillary electrophoresis using on-line concentration method
Analysis of glycoprotein-derived oligosaccharides in glycoproteins detected on two-dimensional gel by capillary electrophoresis using on-line concentration method
Engineering Village 2
? 2006 Elsevier Inc
Accession number: 06059674461
Title: Analysis of glycoprotein-derived oligosaccharides in glycoproteins detected on two-dimensional gel by capillary electrophoresis using on-line concentration method
Authors: Kamoda, Satoru; Nakanishi, Yasuharu; Kinoshita, Mitsuhiro; Ishikawa, Rika; Kakehi, Kazuaki
Author affiliation: Faculty of Pharmaceutical Sciences, Kinki University, Higashi-osaka 577-8502, Japan
Serial title: Journal of Chromatography A
Abbreviated serial title: J. Chromatogr. A
Volume: v 1106
Issue: n 1-2
Monograph title: Microseparation Sciences: Perspectives and Recent Advances with Recognition of Shigeru Terabe
Issue date: Feb 17 2006
Publication year: 2006
Pages: p 67-74
Language: English
ISSN: 0021-9673
CODEN: JCRAEY
Document type: Journal article (JA)
Publisher: Elsevier, Amsterdam, 1000 AE, Netherlands
Abstract: Capillary electrophoresis (CE) is an effective tool to analyze carbohydrate mixture derived from glycoproteins with high resolution. However, CE has a disadvantage that a few nanoliters of a sample solution are injected to a narrow capillary. Therefore, we have to prepare a sample solution of high concentration for CE analysis. In the present study, we applied head column field-amplified sample stacking method to the analysis of N-linked oligosaccharides derived from glycoprotein separated by two-dimensional gel electrophoresis. Model studies demonstrated that we achieved 60-360 times concentration effect on the analysis of carbohydrate chains labeled with 3-aminobenzoic acid (3-AA). The method was applied to the analysis of N-linked oligosaccharides from glycoproteins separated and detected on PAGE gel. Heterogeneity of a1-acid glycoprotein (AGP), i.e. glycoforms, was examined by 2D-PAGE and N-linked oligosaccharides were released by in-gel digestion with PNGase F. The released oligosaccharides were derivatized with 3-AA and analyzed by CE. The results showed that glycoforms having lower pI values contained a larger amount of tetra- and tri-antennary oligosaccharides. In contrast, glycoforms having higher pI values contained bi-antennary oligosaccharides abundantly. The result clearly indicated that the spot of a glycoprotein glycoform detected by Coomassie brilliant blue staining on 2D-PAGE gel is sufficient for quantitative profiling of oligosaccharides. ? 2005 Elsevier B.V. All rights reserved.
Number of references: 41
Ei main heading: Proteins
Ei controlled terms: Glycols - Oligomers - Electrophoresis - Gels - Chemical analysis - Mathematical models
Uncontrolled terms: Capillary electrophoresis - On-line concentration - Glycoproteins - Two-dimensional gel electrophoresis - Oligosaccharides
Ei classification codes: 804.1 Organic Compounds - 815.1.1 Organic Polymers - 701.1 Electricity: Basic Concepts & Phenomena - 801.3 Colloid Chemistry - 801.4.1 Electrochemistry - 921 Applied Mathematics
Treatment: Theoretical (THR); Experimental (EXP)
DOI: 10.1016/j.chroma.2005.08.052
Database: Compendex
Compilation and indexing terms, ? 2006 Elsevier Inc. All rights reserved
This is a subscription site. You will need to subscribe to Engineering Village 2 to view the details.
Votes:6
Comments: 0