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Methods. 2011 Aug;54(4):361-9. doi: 10.1016/j.ymeth.2011.03.004. Epub 2011 Mar 21.

Quantitative profiling of serum samples using TMT protein labelling, fractionation and LC-MS/MS.

Author information

1
Cancer Proteomics Laboratory, EGA Institute for Women's Health, University College London UCL, Cruciform Building, Gower St., London WC1E 6BT, UK.

Abstract

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.

PMID:
21397697
DOI:
10.1016/j.ymeth.2011.03.004
[Indexed for MEDLINE]

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