Capillary Electrophoresis of Sialic Acid-Containing Glycoprotein. Effect of the Heterogeneity of Carbohydrate Chains on Glycoform Separation Using an α1-Acid Glycoprotein as a Model
- 27 April 2001
- journal article
- research article
- Published by American Chemical Society (ACS) in Analytical Chemistry
- Vol. 73 (11) , 2640-2647
- https://doi.org/10.1021/ac001382u
Abstract
α1-Acid glycoprotein (AGP) showed multiple peaks on separation using capillary electrophoresis in a chemically modified capillary with dimethylpolysiloxane at slightly acidic conditions. We analyzed glycoforms of AGP species after separation by ion-exchange chromatography, Con A affinity chromatography, and Cu(II)-chelating affinity chromatography. The AGP species thus obtained were digested with N-glycosidase F, and the released carbohydrate chains were analyzed by high-performance liquid chromatography after labeling with 3-aminobenzoic acid. The results afforded basic information on the contribution of carbohydrate chains to the separation mechanism of glycoforms of AGP by capillary electrophoresis. In addition, we describe an easy method for AGP analysis in serum samples using the electrokinetic injection.Keywords
This publication has 23 references indexed in Scilit:
- Glycosylation of .ALPHA.1-Acid Glycoprotein(Orosomucoid) in Health and Disease: Occurrence, Regulation and Possible Funtional Implications.Trends in Glycoscience and Glycotechnology, 1998
- Analysis of Acidic Oligosaccharides and Glycopeptides by Matrix-Assisted Laser Desorption/Ionization Time-of-Flight Mass SpectrometryAnalytical Chemistry, 1996
- Characterization of changes in the glycosylation pattern of recombinant proteins from BHK-21 cells due to different culture conditionsJournal of Biotechnology, 1995
- Inflammation-induced expression of sialyl Lewis X-containing glycan structures on alpha 1-acid glycoprotein (orosomucoid) in human sera.The Journal of Experimental Medicine, 1993
- Zone broadening in electrophoresis with special reference to high‐performance electrophoresis in capillaries: An interplay between theory and practiceElectrophoresis, 1990
- Effects of N-glycosylation on in vitro activity of Bowes melanoma and human colon fibroblast-derived tissue plasminogen activatorBiochemistry, 1989
- Glycosylation of three molecular forms of human α1‐acid glycoprotein having different interactions with concanavalin AEuropean Journal of Biochemistry, 1988
- Microheterogeneity forms of alpha1-acid glycoprotein as indicators of rheumatoid arthritis activityClinica Chimica Acta; International Journal of Clinical Chemistry, 1987
- Determination of the primary structures of 16 asialo-carbohydrate units derived from human plasma α1-acid glycoprotein by 360-MHz proton NMR spectroscopy and permethylation analysisBiochemistry, 1978
- SEQUENCE COMPARISON OF HUMAN PLASMA α1‐ACID GLYCOPROTEIN AND THE IMMUNOGLOBULINSInternational Journal of Peptide and Protein Research, 1978