Separation of branched sialylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection.
Separation of branched sialylated oligosaccharides using high-pH anion-exchange chromatography with pulsed amperometric detection.
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DOI：
10.1016/0003-2697(89)90708-2
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发表时间：
1989-10
影响因子：
2.9
通讯作者：
Raymond R. Townsend;M. Hardy;Dale A. Cumming;J. P. Carver;Brad Bendiak
中科院分区：
文献类型：
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作者：
Raymond R. Townsend;M. Hardy;Dale A. Cumming;J. P. Carver;Brad Bendiak
用高效阴离子交换层析和脉冲安培检测对来自牛胎蛋白的10种唾液酸寡糖(B.Bendiak，M.Harris-Brandts，S.W.Michnick，J.P.Carver和D.A.Cumming，生物化学；D.A.Cumming，C.G.Hellerqvist，M.Harris-Brandts，S.W.Michnick，J.P.Carver，和B.Bendiak，生物化学，在出版社)进行了层析。在近中性的pH值下，寡糖根据唾液酸带的正负电数而被分离出来；而在碱性pH下，由于氧离子的形成，中性部分的寡糖提高了拆分能力。具体地说，含有Gal-β(1，3)GlcNAc序列的三聚三天线寡糖被更好地保留，并且可以与只含有Gal-β(1，4)GlcNAc单元的低聚糖完全分离。根据α(2，6)和α(2，3)连接的唾液酸(α(2，6)连接的唾液酸减少保留时间)的组合来分离含有相同数量唾液酸量的寡糖。二、三、四和五解寡糖的相对摩尔电化学反应相似(相对于葡萄糖为4.8±14%)。对每个特征寡糖和从胎蛋白释放的寡糖与N-葡聚糖酶的混合物进行洗脱研究。不同来源的牛胎蛋白中唾液酸寡糖的主要种类(双、三、四、五)的相对比例差异很大。
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.