Fractionation and characterization of sialyl linkage isomers of serum N-glycans by CE-MS.
Fractionation and characterization of sialyl linkage isomers of serum N-glycans by CE-MS.
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DOI：
10.1002/jssc.202200223
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发表时间：
2022-09
影响因子：
3.1
通讯作者：
Jacobson, Stephen C.
中科院分区：
文献类型：
--
作者：
Zhou, Xiaomei;Song, Woran;Novotny, Milos V.;Jacobson, Stephen C.
唾液化n -聚糖的结构异构体有助于n -聚糖的多样性和一系列的生物学功能。通过两步唾液酸键特异性烷基酰胺化反应生成的α2,3-键和α2,6键之间的质量差异，可以很容易地通过质谱法区分唾液酸键异构体。为了提高从复杂混合物中识别n -聚糖的能力，我们在第一步烷基酰胺化后增加了一个脱乙酰化步骤，使带负电荷的羧酸在α2,3-唾液酸上再生。与α2,3-唾液酸的n-聚糖异构体在第二步烷基酰胺化之前通过离子交换色谱分离。采用这种改进的烷基酰胺化方法，对唾液化n -聚糖进行富集和稳定，并通过毛细管电泳-质谱和串联质谱进行结构表征。我们在人血清中鉴定了52个唾液化的n -聚糖结构，其中包括107个链异构体，并证实了特定唾液链异构体的位置异构体的存在。由于离子交换分离和CE分离后样品的复杂性降低，n-聚糖的亚结构特征在人血清中被快速评估，并包括相应的核心和天线聚焦和聚乳糖胺的诊断离子。
Structural isomers of sialylated N-glycans contribute to the diversity of the N-glycome and to a range of biological functions. Sialyl linkage isomers can be readily distinguished by mass spectrometry with mass differences between α2,3- and α2,6-linkages generated by a two-step sialic acid linkage-specific alkylamidation. To improve identification of N-glycans from complex mixtures, we added a delactonization step after the first alkylamidation step, which regenerates negatively charged carboxylic acids on α2,3-sialic acids. N-glycan isomers with α2,3-sialic acids are then fractionated by ion-exchange chromatography prior to the second alkylamidation step. With this modified alkylamidation method, sialylated N-glycans were enriched and stabilized for structural characterization by capillary electrophoresis-mass spectrometry and tandem mass spectrometry. We identified 52 sialylated N-glycan structures, including 107 linkage isomers, in human serum and confirmed the presence of positional isomers of specific sialyl linkage isomers. Due to the reduced sample complexity after the ion-exchange fractionation and CE separation, substructural features of N-glycans were rapidly evaluated in human serum and included diagnostic ions corresponding to core- and antenna-fucosylation and poly-lactosamine.
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影响因子：
7.4
作者：
Gaunitz S;Nagy G;Pohl NL;Novotny MV
通讯作者：
Novotny MV
影响因子：
7.4
作者：
Alley, William R., Jr.;Madera, Milan;Mechref, Yehia;Novotny, Milos V.
通讯作者：
Novotny, Milos V.
影响因子：
7.4
作者：
Khatri K;Klein JA;Haserick JR;Leon DR;Costello CE;McComb ME;Zaia J
通讯作者：
Zaia J
影响因子：
4.4
作者：
Biskup, Karina;Braicu, Elena I.;Blanchard, Veronique
通讯作者：
Blanchard, Veronique
影响因子：
7.4
作者：
Ni, Wenqin;Bones, Jonathan;Karger, Barry L.
通讯作者：
Karger, Barry L.