Employing TMT Quantification in a Shotgun-MS Platform
Employing TMT Quantification in a Shotgun-MS Platform
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DOI：
10.1007/978-1-4939-0685-7_12
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发表时间：
2014-01-01
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影响因子：
--
通讯作者：
Timms, John F.
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文献类型：
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作者：
O'Brien, Darragh P.;Timms, John F.
血清蛋白质组可能是疾病生物标志物的理想来源，尽管其复杂性需要新的策略来丰富和量化具有生物标志物效用的低丰度蛋白质。本研究使用免疫消耗、多凝集素分离和肽串联质量标签（TMT）标记的工作流程对诊断前胰腺癌患者和对照组的血清样本进行了比较。然后将样品进行SCX和高pH反相分离，并进行液相色谱-串联质谱（LC-MS/MS）分析。目的是发现胰腺癌的候选血清生物标志物，尽管该方法适用于任何血清样本的比较蛋白质组学分析。
The blood serum proteome may be an ideal source of disease biomarkers, although its complexity necessitates novel strategies to enrich and quantify lower-abundance proteins with biomarker utility. Herein, serum samples from pre-diagnosis pancreatic cancer cases and controls were compared using a workflow of immunodepletion, multi-lectin fractionation, and peptide tandem mass tag (TMT) labeling. Samples were then subjected to SCX and high pH reversed-phase fractionation and analyzed by liquid chromatography tandem mass spectrometry (LC-MS/MS). The aim was the discovery of candidate serum biomarkers of pancreatic cancer, although the method is applicable to any comparative proteomic analysis of serum samples.