Enrichment and identification of glycoproteins in human saliva using lectin magnetic bead arrays
Enrichment and identification of glycoproteins in human saliva using lectin magnetic bead arrays
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DOI：
10.1016/j.ab.2015.11.024
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发表时间：
2016-03-15
影响因子：
2.9
通讯作者：
Punyadeera, Chamindie
中科院分区：
文献类型：
--
作者：
Caragata, Michael;Shah, Alok K.;Punyadeera, Chamindie
蛋白质的异常糖基化是肿瘤发生的标志，并可在癌症检测中提供诊断价值。由于唾液蛋白质组中糖基化蛋白的比例相对较高，人唾液是糖蛋白的理想来源。此外，唾液收集是非侵入性的，技术上简单，样本收集和储存相对容易。尽管蛋白质的差异糖基化可以指示疾病状态，但从临床样品中鉴定差异糖基化并非微不足道。为了促进唾液糖蛋白生物标志物的发现，我们优化了一种基于凝集素磁珠阵列（saLeMBA）的人唾液差异糖蛋白富集方法。在saLeMBA平台中使用来自不同反应性组的选择的凝集素来富集来自健康志愿者唾液的唾液糖蛋白。采用液相色谱串联质谱法（LC-MS/MS）分析saLeMBA的技术重现性，以鉴定每种凝集素富集的糖基化蛋白。我们的saLeMBA平台能够以糖蛋白和凝集素特异性方式实现稳健的糖蛋白富集，与已知的蛋白特异性聚糖谱一致。我们证明了saLeMBA是一种可靠的方法来丰富和检测存在于人唾液中的糖蛋白。(C)2015 Elsevier Inc. All rights reserved.
Aberrant glycosylation of proteins is a hallmark of tumorigenesis and could provide diagnostic value in cancer detection. Human saliva is an ideal source of glycoproteins due to the relatively high proportion of glycosylated proteins in the salivary proteome. Moreover, saliva collection is noninvasive and technically straightforward, and the sample collection and storage is relatively easy. Although differential glycosylation of proteins can be indicative of disease states, identification of differential glycosylation from clinical samples is not trivial. To facilitate salivary glycoprotein biomarker discovery, we optimized a method for differential glycoprotein enrichment from human saliva based on lectin magnetic bead arrays (saLeMBA). Selected lectin from distinct reactivity groups were used in the saLeMBA platform to enrich salivary glycoproteins from healthy volunteer saliva. The technical reproducibility of saLeMBA was analyzed with liquid chromatography tandem mass spectrometry (LC-MS/MS) to identify the glycosylated proteins enriched by each lectin. Our saLeMBA platform enabled robust glycoprotein enrichment in a glycoproteinand lectin-specific manner consistent with known protein-specific glycan profiles. We demonstrated that saLeMBA is a reliable method to enrich and detect glycoproteins present in human saliva. (C) 2015 Elsevier Inc. All rights reserved.