A general approach to desalting oligosaccharides released from glycoproteins
A general approach to desalting oligosaccharides released from glycoproteins
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DOI：
10.1023/a:1006983125913
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发表时间：
1998-08-01
影响因子：
3
通讯作者：
Redmond, JW
中科院分区：
文献类型：
--
作者：
Packer, NH;Lawson, MA;Redmond, JW
糖样品的脱盐对于许多碳水化合物分析技术的成功是必不可少的，例如质谱法、毛细管电泳、阴离子交换色谱、酶降解和化学衍生。目前使用的所有分离方法都有局限性：例如，混合床离子交换柱有损失带电糖的风险，通过非水溶剂沉淀盐可能导致寡糖共沉淀，凝胶色谱法使用高度交联的填料，其中难以实现小寡糖的分离。我们证明石墨化碳作为固相萃取柱可用于寡糖的纯化（或其衍生物）从含有以下一种或多种污染物的溶液中分离：盐（包括氢氧化物、乙酸盐、磷酸盐）、单糖、洗涤剂（十二烷基硫酸钠和Triton X-100），蛋白质（包括酶）和从糖缀合物中释放寡糖的试剂（如肼和硼氢化钠）。从吸附剂中完全回收寡糖，其也可用于分离酸性和中性聚糖。具体的应用，如清理N-连接的寡糖后，通过PNGase F和肼，O-连接的聚糖的去除后，通过碱，在线的HPAEC分离的寡糖和β-消除糖醇的电喷雾质谱法之前，从尿液中的寡糖纯化。
Desalting of sugar samples is essential for the success of many techniques of carbohydrate analysis such as mass spectrometry, capillary electrophoresis, anion exchange chromatography, enzyme degradation and chemical derivatization. All desalting methods which are currently used have limitations: for example, mixed-bed ion-exchange columns risk the loss of charged sugars, precipitation of salt by a non-aqueous solvent can result in co-precipitation of oligosaccharides, and gel chromatography uses highly crosslinked packings in which separation of small oligosaccharides is difficult to achieve. We demonstrate that graphitized carbon as a solid phase extraction cartridge can be used for the purification of oligosaccharides (or their derivatives) from solutions containing one or more of the following contaminants: salts (including salts of hydroxide, acetate, phosphate), monosaccharides, detergents (sodium dodecyl sulfate and Triton X-100), protein (including enzymes) and reagents for the release of oligosaccharides from glycoconjugates (such as hydrazine and sodium borohydride). There is complete recovery of the oligosaccharides from the adsorbent which can also be used to fractionate acidic and neutral glycans. Specific applications such as clean-up of N-linked oligosaccharides after removal by PNGase F and hydrazine, desalting of O-linked glycans after removal by alkali, on-line desalting of HPAEC-separated oligosaccharides and beta-eliminated alditols prior to electrospray mass spectrometry, and purification of oligosaccharides from urine are described.