title: Protein profile of exhaled breath condensate determined by high resolution mass spectrometry.
abstract: A method based on liquid chromatography/high resolution tandem mass spectrometry coupled with electrophoretic separation, for determination and relative quantification of the protein composition of exhaled breath condensate (EBC), was developed. Application of the procedure to a sample of EBC, pooled from nine healthy subjects, resulted in the identification of 167 unique gene products, 113 of which not previously reported in EBC samples. The abundance of the protein identified was estimated by means of the exponentially modified protein abundance index protocol (emPAI). Cytokeratins were by far the most abundant proteins in EBC samples. Many of the identified proteins were associated with multiple cellular location with cytoplasm constituting the largest group. Cytosol, nucleus, membrane, cytoskeleton and extracellular were other abundantly represented locations. No amylase was detected, suggesting the absence of saliva protein contamination. The profile obtained represents the most comprehensive protein characterization of EBC so far reported and demonstrates that this approach provides a powerful tool for investigating the protein profile of EBC samples. Compared with analogous investigations, this study also shows that the protein profile of EBC is strongly affected by the sampling method adopted.
authors.affiliation: Department of Chemical Sciences, University of Catania, Viale A. Doria 6, 95125 Catania, Italy.
authors.forename: Vera
authors.initials: V
authors.lastname: Muccilli
authors.affiliation: Department of Chemical Sciences, University of Catania, Viale A. Doria 6, 95125 Catania, Italy. Electronic address: rsaletti@unict.it.
authors.forename: Rosaria
authors.initials: R
authors.lastname: Saletti
authors.affiliation: Department of Chemical Sciences, University of Catania, Viale A. Doria 6, 95125 Catania, Italy.
authors.forename: Vincenzo
authors.initials: V
authors.lastname: Cunsolo
authors.affiliation: ThermoFisher Scientific, Stafford House, Boundary Way, Hemel Hempstead HP2 7GE, United Kingdom.
authors.forename: Jenny
authors.initials: J
authors.lastname: Ho
authors.affiliation: Department of Clinical and Molecular Biomedicine, University Hospital Policlinico-Vittorio Emanuele, Via Santa Sofia 78, 95123 Catania, Italy.
authors.forename: Elisa
authors.initials: E
authors.lastname: Gili
authors.affiliation: Department of Clinical and Molecular Biomedicine, University Hospital Policlinico-Vittorio Emanuele, Via Santa Sofia 78, 95123 Catania, Italy.
authors.forename: Enrico
authors.initials: E
authors.lastname: Conte
authors.affiliation: Department of Clinical and Molecular Biomedicine, University Hospital Policlinico-Vittorio Emanuele, Via Santa Sofia 78, 95123 Catania, Italy.
authors.forename: Stefania
authors.initials: S
authors.lastname: Sichili
authors.affiliation: Department of Clinical and Molecular Biomedicine, University Hospital Policlinico-Vittorio Emanuele, Via Santa Sofia 78, 95123 Catania, Italy.
authors.forename: Carlo
authors.initials: C
authors.lastname: Vancheri
authors.affiliation: Department of Chemical Sciences, University of Catania, Viale A. Doria 6, 95125 Catania, Italy.
authors.forename: Salvatore
authors.initials: S
authors.lastname: Foti
mesh_terms: D000328:Adult; D001944:Breath Tests / Q000295:instrumentation / Q000379:methods*; D002851:Chromatography, High Pressure Liquid / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D008297:Male; D011506:Proteins / Q000302:isolation & purification*; D040901:Proteomics; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D011506:Proteins
keywords: EBC protein profile; Exhaled breath condensate (EBC); LTQ-Orbitrap; Lung disease; Proteomics