Proteomic analysis of undiluted lung epithelial lining fluid.
BACKGROUND
Proteomics is increasingly leading to biomarker discovery in human disease. Epithelial lining fluid (ELF), until now only recovered indirectly, diluted in BAL fluid, is an attractive sample for lung disease proteomics. The direct recovery of undiluted ELF is now possible using a bronchoscopic microsampling (BMS) probe. In this preliminary study of anesthetized ventilated rabbits, we applied this probe to recover ELF and to analyze the resulting samples with the aim of determining their potential in lung disease biomarker discovery.


METHODS
In order to do so, a method was devised and evaluated in preliminary experiments both for nonbronchoscopic use of the probe and for recovering undiluted ELF from probe tips. To verify the proteomic potential of the sample, the recovered ELF was separated by one-dimensional polyacrylamide gel electrophoresis, and the resulting lane was cut into multiple fractions, each of which was digested and analyzed by liquid chromatography tandem mass spectrometry. The identified proteins were then searched against Medline for association with broad categories of lung disease.


RESULTS
Nonbronchoscopic use of the probe allowed successful ELF sampling and the recovery of undiluted ELF from probe tips. Proteomic analysis showed that ELF contains many proteins that have already been reported as being associated with lung disease as well as proteins potentially correlated with lung disease.


CONCLUSIONS
This preliminary study of undiluted ELF, as recovered by the BMS probe, shows that it may be an ideal sample for lung proteomics. The potential application of this sampling technique in various lung diseases will need to be confirmed by future studies.
Kipnis
Eric
E
Infectious Diseases Research Laboratory, Department of Inflammation and Sepsis, Lille University Medical School, Lille, France. Electronic address: ekipnis@gmail.com.
Hansen
Kirk
K
Cancer Center Proteomics Core, University of Colorado Denver, Aurora, CO.
Sawa
Teiji
T
Department of Anesthesia and Perioperative Care, University of California San Francisco, San Francisco, CA.
Moriyama
Kiyoshi
K
Department of Anesthesia and Perioperative Care, University of California San Francisco, San Francisco, CA.
Zurawel
Ashley
A
Cancer Center Proteomics Core, University of Colorado Denver, Aurora, CO.
Ishizaka
Akitoshi
A
Department of Medicine, Keio University, Tokyo, Japan.
Wiener-Kronish
Jeanine
J
Department of Anesthesia and Critical Care, Massachusetts General Hospital, Boston, MA.
D000818:Animals; D001992:Bronchoalveolar Lavage Fluid / Q000737:chemistry*; D020680:Bronchoscopes; D002406:Catheterization, Peripheral / Q000295:instrumentation*; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D005240:Feasibility Studies; D008171:Lung Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D008297:Male; D040901:Proteomics; D011817:Rabbits; D020545:Respiratory Mucosa / Q000737:chemistry*; D053719:Tandem Mass Spectrometry