Cancer Proteomics Laboratory, EGA Institute for Women's Health, University College London UCL, Cruciform Building, Gower St., London WC1E 6BT, UK.
Methods. 2011 Aug;54(4):361-9. doi: 10.1016/j.ymeth.2011.03.004. Epub 2011 Mar 21.
Blood-borne biomarkers are urgently required for the early detection, accurate diagnosis and prognosis of disease. Additionally, improved methods of profiling serum and plasma proteins for biomarker discovery efforts are needed. Herein, we report a quantitative method based on amino-group labelling of serum proteins (rather than peptides) with isobaric tandem mass tags (TMT) and incorporating immune-based depletion, gel-based and strong anion exchange separation of proteins prior to differential endoproteinase treatment and liquid chromatography tandem mass spectrometry. We report a generally higher level of quantitative coverage of the serum proteome compared to other peptide-based isobaric tagging approaches and show the potential of the method by applying it to a set of unique samples that pre-date the diagnosis of pancreatic cancer.
目前非常需要血液生物标志物来进行疾病的早期检测、准确诊断和预后判断。此外，还需要改进用于生物标志物发现的血清和血浆蛋白质分析方法。在此，我们报告了一种基于血清蛋白质（而非肽段）的氨基标记的定量方法，该方法使用了同位素质谱标签（TMT），并结合了免疫清除、凝胶电泳和强阴离子交换分离蛋白质，然后进行差异内切蛋白酶处理和液相色谱串联质谱分析。与其他基于肽段的同位素质谱标签方法相比，我们报告了更高水平的血清蛋白质组定量覆盖度，并通过应用于一组独特的、在胰腺癌诊断之前的样本中展示了该方法的潜力。
