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使用血清素键合硅胶特异性提取含唾液酸聚糖和糖肽。

Specific extraction of sialic-acid-containing glycans and glycopeptides using serotonin-bonded silica.

作者信息

Yodoshi Masahiro, Ikuta Tomoko, Mouri Yukie, Suzuki Shigeo

机构信息

Department of Pharmaceutical Sciences, Kinki University, 3-4-1 Kowakae, Higashi-osaka, Osaka 577-8502, Japan.

出版信息

Anal Sci. 2010 Jan;26(1):75-81. doi: 10.2116/analsci.26.75.

Abstract

Serotonin-bonded silica was developed for specific extraction of sialic-acid-containing glycans and glycopeptides. Serotonin, coupled by reductive amination with aldehyde silica particles via its ethylamino group, has strong affinity to sialic acid in glycan chain termini. Sialylated glycans trapped on serotonin silica particles are released by washing with ammonium acetate solution, providing highly efficient specific trap and release glycoconjugates. With >100 micromol/g adsorption capacity, the particles are applicable to purify labeled glycans after derivatization with 2-aminopyridine to remove excess reagents. Serotonin silica efficiently enriches sialic-acid-containing glycopeptides from tryptic digests for LC/MS analysis of glycans' heterogeneity in glycoproteins.

摘要

血清素键合硅胶被开发用于特异性提取含唾液酸的聚糖和糖肽。血清素通过其乙氨基与醛基硅胶颗粒进行还原胺化反应而偶联,对聚糖链末端的唾液酸具有很强的亲和力。捕获在血清素硅胶颗粒上的唾液酸化聚糖通过用醋酸铵溶液洗涤而释放,从而实现高效的特异性捕获和释放糖缀合物。该颗粒具有大于100微摩尔/克的吸附容量,适用于在用2-氨基吡啶衍生化以去除过量试剂后纯化标记的聚糖。血清素硅胶能有效地从胰蛋白酶消化物中富集含唾液酸的糖肽,用于糖蛋白中聚糖异质性的液相色谱/质谱分析。

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