The INLIGHT™ strategy for N-linked glycan derivatization has been shown to overcome many of the challenges associated with glycan analysis. The hydrazide tag reacts efficiently with the glycans, increasing their non-polar surface area, allowing for reversed-phase separations and increased ionization efficiency. We have taken the INLIGHT™ strategy and adopted it for use with O-linked glycans. A central composite design was utilized to find optimized tagging conditions (45% acetic acid, 0.1 μg/μL tag concentration, 37 C, 1.75 h). Derivatization at optimized conditions was much quicker than any hydrazide derivatization strategy used previously. Human immunoglobulin A (IgA) and bovine submaxillary mucin (BSM) were then deglycosylated through hydrazinolysis and the removed glycans were tagged under optimum conditions. XIC of tagged glycans and MS2 data show successful hydrazide tagging of O-linked glycans for the first time.

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N联聚糖衍生化的INLIGHT™策略已被证明克服了与聚糖分析相关的许多挑战。酰肼标签与聚糖有效反应，增加了它们的非极性表面积，可以进行反相分离并提高电离效率。我们采用了INLIGHT™策略，并将其用于O链聚糖。利用中央复合材料设计来找到优化的标记条件（45％乙酸，0.1μg/μL标记浓度，37°C，1.75 h）。在优化条件下进行衍生化要比以前使用的任何酰肼衍生化策略快得多。然后通过肼解作用将人免疫球蛋白A（IgA）和牛上颌黏液（BSM）糖基化，并在最佳条件下标记去除的聚糖。标记的聚糖的XIC和MS2数据首次显示O-连接的聚糖成功进行酰肼标记。

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