title: Proteomic analysis of human epithelial lining fluid by microfluidics-based nanoLC-MS/MS: a feasibility study.
abstract: Microfluidics-based nanoLC-MS/MS (chipLC-MS/MS) was used to identify and quantify proteins in epithelial lining fluid (ELF), collected during bronchoscopy from the main bronchi of chronic obstructive pulmonary disease (COPD) patients and healthy controls using microprobes. ELF is a biofluid that is well suited to study pathophysiological processes in the lung, because it contains high concentrations of biologically active molecules. 1D-PAGE followed by in-gel tryptic digestion and chipLC-MS/MS resulted in identification of approximately 300 proteins. A comparative study of ELF from COPD patients and non-COPD controls using chemical stable isotope labeling (iTRAQ®-8Plex) showed that the levels of lactotransferrin, high-mobility group protein B1 (HMGB 1), alpha 1-antichymotrypsin and cofilin-1 differed significantly in ELF from COPD patients and non-COPD controls (p-values < 0.05). These results were reproduced in another, independent set of ELF samples from COPD patients and non-COPD controls and further validated by immunohistochemistry. This study shows the feasibility of performing chipLC-MS/MS and quantitative proteomics in human ELF.
authors.forename: Lorenza
authors.initials: L
authors.lastname: Franciosi
authors.forename: Natalia
authors.initials: N
authors.lastname: Govorukhina
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Fusetti
authors.forename: Bert
authors.initials: B
authors.lastname: Poolman
authors.forename: Monique E
authors.initials: ME
authors.lastname: Lodewijk
authors.forename: Wim
authors.initials: W
authors.lastname: Timens
authors.forename: Dirkje
authors.initials: D
authors.lastname: Postma
authors.forename: Nick
authors.initials: N
authors.lastname: ten Hacken
authors.forename: Rainer
authors.initials: R
authors.lastname: Bischoff
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001826:Body Fluids / Q000737:chemistry*; D016022:Case-Control Studies; D002853:Chromatography, Liquid / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D005260:Female; D006801:Humans; D007150:Immunohistochemistry; D008168:Lung / Q000737:chemistry*; D008297:Male; D046210:Microfluidic Analytical Techniques / Q000379:methods*; D008875:Middle Aged; D036103:Nanotechnology / Q000379:methods; D010446:Peptide Fragments / Q000032:analysis / Q000737:chemistry; D020543:Proteome / Q000032:analysis* / Q000737:chemistry; D040901:Proteomics; D029424:Pulmonary Disease, Chronic Obstructive / Q000378:metabolism; D015203:Reproducibility of Results; D020545:Respiratory Mucosa / Q000737:chemistry*; D053719:Tandem Mass Spectrometry / Q000379:methods
chemical_list: D010446:Peptide Fragments; D020543:Proteome