The study of carbohydrates requires large amounts of glycans. N-Glycans can be synthesized but generating large quantities of N-glycans with diverse structures remains difficult. In this study, we aimed to obtain large amounts of glycans using an optimized procedure. Two types of reductive N-glycans were released from chicken egg albumin (ovalbumin) and soy protein using an ammonia catalysis method and labeled with benzenesulfonyl hydrazide (BSH). After preliminary separation by preparative HPLC, N-glycan-BSH components were de-labeled separately and reducing N-glycans were recovered. The de-labeled reducing N-glycans were derived with different labeling reagents and further separated and purified with two/multi-dimensional HPLC for various studies. We selected the bifunctional reagent 2-amino-N-(2-aminoethyl)-benzamide (AEAB) as a labeling reagent combined with C18 column for two-dimensional HPLC separation. A total of 21 and 8 N-glycan-AEAB conjugates were obtained from ovalbumin and soy protein, respectively. A reactive primary alkylamine of N-glycan-AEAB conjugates can be effectively immobilized on microarray surfaces, allowing for subsequent functional studies of glycans.

中文翻译：

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基于氨催化释放法的N-聚糖的分离与制备。

碳水化合物的研究需要大量的聚糖。Ñ -Glycans可以合成，但产生大量的Ñ -glycans具有不同的结构仍然是困难的。在这项研究中，我们旨在使用优化的程序获得大量的聚糖。使用氨催化方法从鸡蛋清蛋白（卵清蛋白）和大豆蛋白中释放出两种类型的还原性N-聚糖，并用苯磺酰肼（BSH）标记。通过制备型HPLC初步分离后，分别对N-聚糖-BSH组分进行脱标记，并回收还原性N-聚糖。去标记的还原氮用不同的标记试剂衍生β-聚糖，并进一步分离和纯化，用二维/多维HPLC进行各种研究。我们选择双功能试剂2-氨基-N-（2-氨基乙基）-苯甲酰胺（AEAB）作为标记试剂，并与C18色谱柱组合用于二维HPLC分离。分别从卵清蛋白和大豆蛋白中获得总共21和8个 N-聚糖-AEAB缀合物。N-聚糖-AEAB共轭物的反应性伯烷基胺可以有效地固定在微阵列表面上，从而可以进行聚糖的后续功能研究。