TY - JOUR
T1 - Separation of branched sialylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection
AU - Reid Townsend, R.
AU - Hardy, Mark R.
AU - Cumming, Dale A.
AU - Carver, Jeremy P.
AU - Bendiak, Brad
N1 - Funding Information:
R.R.T. was supported by the National Institutes of Health Grant DK31376. M.R.H. was the recipient of a Postdoctoral Carbohydrate Research Fellowship from Dionex Corporation (Sunnyvale, CA). The authors thank the MRC of Canada and the Terry Fox Special Initiatives Program for support. B.B. thanks Harry Schachter for MRC support. The Dionex carbohydrate analyzer was purchased with an instrumentation supplement grant to National Institutes of Health Research DK-09970 to Dr. Yuan Chuan Lee. This paper is contribution 1483 from the McCollum-Pratt Institute, The Johns Hopkins The authors thank Dr. Chuan Lee for the use of for the chromatography
PY - 1989/10
Y1 - 1989/10
N2 - Ten characterized sialylated oligosaccharides from bovine fetuin (B. Bendiak, M. Harris-Brandts, S. W. Michnick, J. P. Carver, and D. A. Cumming, Biochemistry, in press; and D. A. Cumming, C. G. Hellerqvist, M. Harris-Brandts, S. W. Michnick, J. P. Carver, and B. Bendiak, Biochemistry, in press) were chromatographed using high-performance anion-exchange chromatography with pulsed amperometric detection. At near neutral pH values, oligosaccharides were separated according to their number of formal negative charges from sialic acid; however, at alkaline pH, the neutral portion of the oligosaccharides enhanced resolution due to oxyanion formation. Specifically, trisialylated triantennary oligosaccharides containing a Gal-β(1,3)GlcNAc sequence were more retained and could be completely separated from those having only Gal-β(1,4)GlcNAc units. Oligosaccharides containing the same number of sialic acids were separated according to the combination of α(2,6)- and α(2,3)-linked sialic acids (α(2,6)-linked sialic acid reduced retention time). The relative molar electrochemical responses for di-, tri-, tetra-, and pentasialylated oligosaccharides were found to be similar (4.8 ± 14% relative to glucose). Coelution studies were performed with each of the characterized oligosaccharides and the mixture of oligosaccharides which were released from fetuin with N-glycanase. The relative proportion of the major classes of sialylated oligosaccharides (bi-, tri-, tetra-, and penta-) varied significantly in bovine fetuin from different sources.
AB - Ten characterized sialylated oligosaccharides from bovine fetuin (B. Bendiak, M. Harris-Brandts, S. W. Michnick, J. P. Carver, and D. A. Cumming, Biochemistry, in press; and D. A. Cumming, C. G. Hellerqvist, M. Harris-Brandts, S. W. Michnick, J. P. Carver, and B. Bendiak, Biochemistry, in press) were chromatographed using high-performance anion-exchange chromatography with pulsed amperometric detection. At near neutral pH values, oligosaccharides were separated according to their number of formal negative charges from sialic acid; however, at alkaline pH, the neutral portion of the oligosaccharides enhanced resolution due to oxyanion formation. Specifically, trisialylated triantennary oligosaccharides containing a Gal-β(1,3)GlcNAc sequence were more retained and could be completely separated from those having only Gal-β(1,4)GlcNAc units. Oligosaccharides containing the same number of sialic acids were separated according to the combination of α(2,6)- and α(2,3)-linked sialic acids (α(2,6)-linked sialic acid reduced retention time). The relative molar electrochemical responses for di-, tri-, tetra-, and pentasialylated oligosaccharides were found to be similar (4.8 ± 14% relative to glucose). Coelution studies were performed with each of the characterized oligosaccharides and the mixture of oligosaccharides which were released from fetuin with N-glycanase. The relative proportion of the major classes of sialylated oligosaccharides (bi-, tri-, tetra-, and penta-) varied significantly in bovine fetuin from different sources.
UR - http://www.scopus.com/inward/record.url?scp=0024454323&partnerID=8YFLogxK
U2 - 10.1016/0003-2697(89)90708-2
DO - 10.1016/0003-2697(89)90708-2
M3 - Article
C2 - 2481411
AN - SCOPUS:0024454323
SN - 0003-2697
VL - 182
SP - 1
EP - 8
JO - Analytical Biochemistry
JF - Analytical Biochemistry
IS - 1
ER -