NIBRT Dublin-Oxford Glycobiology Laboratory, NIBRT-The National Institute for Bioprocessing Research and Training, UCD Conway Institute of Biomolecular and Biomedical Research, University College Dublin, Belfield, Dublin 4, Ireland.
Anal Chem. 2010 Dec 15;82(24):10208-15. doi: 10.1021/ac102860w. Epub 2010 Nov 12.
Glycosylation is a diverse but critically important post-translational modification that modulates the physical, chemical and biological properties of proteins. Alterations in glycosylation have been noted in a number of diseases including cancer. The discovery of alterations in the glycosylation of serum glycoproteins which may offer potential as biomarkers is attracting considerable research interest. In the current study, the significant improvements in efficiency, selectivity, and analysis speed offered by ultra performance liquid chromatography (UPLC) profiling of fluorescently labeled N-linked oligosaccharides on a recently introduced sub-2 μm hydrophilic interaction (HILIC) based stationary phase are demonstrated to identify cancer associated alterations in the serum N-glycome of patients bearing stomach adenocarcinoma. The contribution of the glycosylation present on four highly abundant serum proteins namely, IgG, haptoglobin, transferrin, and α1-acid glycoprotein was evaluated. Alterations in the glycosylation present on these four proteins isolated from the pathologically staged cancer serum using either affinity purification or two-dimensional electrophoresis were then investigated as possible markers for stomach cancer progression. In agreement with previous reports, an increase in sialylation was observed on haptoglobin, transferrin, and α1-acid glycoprotein in the cancerous state. Increased levels of core fucosylated biantennary glycans and decreased levels of monogalactosylated core fucosylated biantennary glycans were present on IgG with increasing disease progression. The speed and selectivity offered by the sub-2 μm HILIC phase make it ideal for rapid yet highly efficient separation of complex oligosaccharide mixtures such as that present in the serum N-glycome.
糖基化是一种多样化但至关重要的翻译后修饰，可调节蛋白质的物理、化学和生物学特性。在许多疾病中都观察到糖基化的改变，包括癌症。血清糖蛋白糖基化的改变可能作为生物标志物提供潜在的研究兴趣。在当前的研究中，通过最近推出的基于亚 2μm 亲水性相互作用（HILIC）固定相的荧光标记 N-连接寡糖的超高效液相色谱（UPLC）分析，展示了效率、选择性和分析速度的显著提高，用于鉴定患有胃腺癌的患者血清 N-聚糖中的癌症相关改变。评估了存在于四种高度丰富的血清蛋白（即 IgG、触珠蛋白、转铁蛋白和α1-酸性糖蛋白）上的糖基化的贡献。然后，研究了从病理分期的癌症血清中使用亲和纯化或二维电泳分离出的这四种蛋白质上存在的糖基化的改变是否可能作为胃癌进展的标志物。与先前的报告一致，在癌症状态下观察到触珠蛋白、转铁蛋白和α1-酸性糖蛋白上的唾液酸化增加。随着疾病的进展，在 IgG 上存在更多的核心岩藻糖基化双天线聚糖和更少的单半乳糖基化核心岩藻糖基化双天线聚糖。亚 2μm HILIC 相提供的速度和选择性使其成为快速且高效分离复杂寡糖混合物（如血清 N-聚糖中存在的寡糖混合物）的理想选择。
