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Proteomic analysis of human epithelial lining fluid by microfluidics‐based nano LC ‐ MS / MS : A feasibility study
Author(s) -
Franciosi Lorenza,
Govorukhitalia,
Fusetti Fabrizia,
Poolman Bert,
Lodewijk Monique E.,
Timens Wim,
Postma Dirkje,
ten Hacken Nick,
Bischoff Rainer
Publication year - 2013
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201300020
Subject(s) - chemistry , microfluidics , chromatography , nanotechnology , materials science
Microfluidics‐based nano LC ‐ MS / MS (chip LC ‐ 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 chip LC ‐ 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 (i TRAQ ®‐8 P lex) showed that the levels of lactotransferrin, high‐mobility group protein B 1 ( 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 chip LC ‐ MS / MS and quantitative proteomics in human ELF .