Supported Molecular Matrix Electrophoresis: A New Tool for Characterization of Glycoproteins
Supported Molecular Matrix Electrophoresis: A New Tool for Characterization of Glycoproteins
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DOI：
10.1021/ac900157c
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发表时间：
2009-05-15
影响因子：
7.4
通讯作者：
Kameyama, Akihiko
中科院分区：
文献类型：
--
作者：
Matsuno, Yu-ki;Saito, Takuro;Kameyama, Akihiko
介绍了一种新概念的分离技术--支撑分子基质电泳（SMNIE）.在SMME中，分析物在由骨架材料支撑的分子基质中迁移。在这里，我们介绍了一种新的策略，分离和表征粘蛋白使用SMME。粘蛋白是一种与肿瘤高度相关的糖蛋白，作为临床诊断多种恶性肿瘤的生物标志物具有很大的潜力。然而，由于粘蛋白的大尺寸、聚合性质和异质糖基化，粘蛋白的分析已经落后于现代技术。对于粘蛋白分析，我们采用聚偏二氟乙烯（PVDF）膜和聚乙烯醇（PVA）作为骨架材料和基质分子，分别。结合SMME与质谱和毛细管电泳，我们表明，粗猪胃粘蛋白由中性和硫酸化粘蛋白和硫酸软骨素含蛋白多糖和透明质酸污染。此外，为了证明生物标志物发现策略的可行性，我们分析了人胰液中的粘蛋白，这是胰腺肿瘤临床生物标志物的重要来源。这项工作揭示了人胰液中存在三种类型的粘蛋白，具有不同的聚糖谱。
A new concept of separation technology, supported molecular matrix electrophoresis (SMNIE), is described. In SMME, analytes migrate in a molecular matrix supported by backbone materials. Here we introduce a novel strategy for the separation and characterization of mucins using SMME. Mucin, a highly tumor-associated glycoprotein, has great potential as clinical biomarker for diagnosis of various malignant tumors. However, due to their large size, polymeric nature, and heterogeneous glycosylation, analysis of mucins has been left behind by modem techniques. For mucin analysis, we employed a poly(vinylidene difluoride) (PVDF) membrane and poly(vinyl alcohol) (PVA) as the backbone material and the matrix molecule, respectively. Combining SMME with mass spectrometry and capillary electrophoresis, we demonstrate that a crude porcine stomach mucin consists of a neutral and a sulfated mucin and is contaminated by chondroitin sulfate-containing proteoglycan and hyaluronic acid. Furthermore, to demonstrate the feasibility of the strategy for biomarker discovery, we analyzed mucins in human pancreatic juice, which is an important source for clinical biomarkers of pancreatic tumors. This work revealed the presence of three types of mucin with distinct glycan profiles in human pancreatic juice.