Abstract
In the search for novel biomarkers, noncandidate-based proteomic strategies open up new opportunities to gain a deeper insight into disease processes regarding their molecular mechanisms, the risk factors involved, and the monitoring of disease progression. To carry out these complex analyses, the combined use of gel electrophoresis with mass spectrometry (MS) represents a powerful choice. In addition, the introduction of protein dye labeling has notably improved the reliability of differential expression studies by increasing the statistical significance of the protein candidates. Here, we describe a strategy where different layers (luminal/abluminal) from the intraluminal thrombus (ILT) of human abdominal aortic aneurysm (AAA) patients were incubated in protein-free medium. Then, the levels of the proteins released were compared by two-dimensional differential in-gel electrophoresis (2D-DIGE) and the proteins of interest identified by MS. We consider that the use of tissue-conditioned media could offer a substantial advantage in the analytical study of biological fluids, as they provide a source of proteins to be released to the bloodstream, which could serve as potential circulating biomarkers.
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References
Blanco-Colio LM et al (2009) Vascular proteomics, a translational approach: from traditional to novel proteomic techniques. Expert Rev Proteomics 6:461–464
Nordon I et al (2009) The role of proteomic research in vascular disease. J Vasc Surg 49:1602–1612
Martinez-Pinna R et al (2008) Proteomics in atherosclerosis. Curr Atheroscler Rep 10:209–215
Martinez-Pinna R et al (2011) Identification of peroxiredoxin-1 as a novel biomarker of abdominal aortic aneurysm. Arterioscler Thromb Vasc Biol 31:935–943
Shevchenko A et al (2006) In-gel digestion for mass spectrometric characterization of proteins and proteomes. Nat Protoc 1:2856–2860
Schuerenberg M et al (2000) Prestructured MALDI-MS sample supports. Anal Chem 72:3436–3442
Suckau D et al (2003) A novel MALDI LIFT-TOF/TOF mass spectrometer for proteomics. Anal Bioanal Chem 376:952–965
Perkins DN et al (1999) Probability-based protein identification by searching sequence databases using mass spectrometry data. Electrophoresis 20:3551–3567
Acknowledgments
This chapter has been supported by the EC, FAD project (FP-7, HEALTH F2-2008-200647), the Spanish MICIN (SAF2010/21852), Ministerio de Sanidad y Consumo, Instituto de Salud Carlos III, Redes RECAVA (RD06/0014/0035), EUS2008-03565, and Fundacion Pro CNIC.
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Martinez-Pinna, R. et al. (2013). Identification of Novel Biomarkers of Abdominal Aortic Aneurysms by 2D-DIGE and MALDI-MS from AAA-Thrombus-Conditioned Media. In: Vivanco, F. (eds) Vascular Proteomics. Methods in Molecular Biology, vol 1000. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-405-0_8
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DOI: https://doi.org/10.1007/978-1-62703-405-0_8
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Publisher Name: Humana Press, Totowa, NJ
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