title: Identification of novel biomarkers of abdominal aortic aneurysms by 2D-DIGE and MALDI-MS from AAA-thrombus-conditioned media.
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.
authors.affiliation: Vascular Research Lab IIS-Fundación Jiménez Díaz, Instituto de Investigacion Sanitaria Fundacion Jimenez Diaz (IIS-FJD), Universidad Autónoma de Madrid, Madrid, Spain.
authors.forename: Roxana
authors.initials: R
authors.lastname: Martinez-Pinna
authors.forename: Juan Antonio
authors.initials: JA
authors.lastname: Lopez
authors.forename: Priscila
authors.initials: P
authors.lastname: Ramos-Mozo
authors.forename: Luis M
authors.initials: LM
authors.lastname: Blanco-Colio
authors.forename: Emilio
authors.initials: E
authors.lastname: Camafeita
authors.forename: Enrique
authors.initials: E
authors.lastname: Calvo
authors.forename: Olivier
authors.initials: O
authors.lastname: Meilhac
authors.forename: Jean Baptiste
authors.initials: JB
authors.lastname: Michel
authors.forename: Jesús
authors.initials: J
authors.lastname: Egido
authors.forename: José Luis
authors.initials: JL
authors.lastname: Martin-Ventura
mesh_terms: D017544:Aortic Aneurysm, Abdominal / Q000150:complications* / Q000378:metabolism*; D015415:Biomarkers / Q000378:metabolism; D017077:Culture Media, Conditioned; D057225:Data Mining; D030562:Databases, Protein; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D059748:Proteolysis; D040901:Proteomics / Q000379:methods*; D016622:Silver Staining; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / Q000379:methods*; D013927:Thrombosis / Q000150:complications / Q000473:pathology*; D014357:Trypsin / Q000378:metabolism
chemical_list: D015415:Biomarkers; D017077:Culture Media, Conditioned; D014357:Trypsin