Identification of pancreas-specific proteins in endoscopically (endoscopic pancreatic function test) collected pancreatic fluid with liquid chromatography--tandem mass spectrometry

Pancreas. 2010 Aug;39(6):889-96. doi: 10.1097/MPA.0b013e3181cf16f4.

Abstract

Objectives: We aimed to establish the endoscopic pancreatic function test (ePFT) as a method that can safely obtain pancreatic fluid for mass spectrometric analysis from patients during upper endoscopy and to reproducibly identify pancreas-specific proteins.

Methods: We performed a sodium dodecyl sulfate-polyacrylamide gel electrophoresis and mass spectrometry-based proteomic analysis (in-gel tryptic digestion followed by liquid chromatography-tandem mass spectrometry [GeLC-MS/MS]) on ePFT-collected pancreatic fluid from 3 individuals, without evidence of chronic pancreatitis, who were undergoing an upper endoscopy for dyspepsia and chronic abdominal pain.

Results: Pancreatic fluid was safely collected from all subjects. The sodium dodecyl sulfate-polyacrylamide gel electrophoretic analysis of ePFT-collected pancreatic fluid revealed no significant variation (F statistic, 1.33, P = 0.29) in protein concentration during the 1-hour collection period and a visually reproducible protein banding pattern among the 3 subjects. The GeLC-MS/MS analysis of ePFT-collected fluid identified pancreas-specific proteins previously described from endoscopic retrograde cholangiopancreatography and surgical collection methods. Gene ontology further revealed that most of the proteins identified have a molecular function of proteases.

Conclusions: The ePFT is capable of collecting large amounts of pancreatic fluid for proteomic analysis enabling the identification of pancreas-specific proteins. This endoscopic collection method coupled with GeLC-MS/MS is a powerful technique, which can be used in future investigations to elucidate pathways involved in the development and progression of pancreatic disease.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Body Fluids / metabolism
  • Chromatography, Liquid / methods*
  • Cohort Studies
  • Electrophoresis, Polyacrylamide Gel
  • Endoscopy, Digestive System
  • Female
  • Humans
  • Male
  • Middle Aged
  • Pancreas / metabolism*
  • Pancreas / pathology
  • Pancreas / physiopathology
  • Pancreatic Function Tests / methods
  • Proteins / analysis*
  • Proteomics / methods
  • Reproducibility of Results
  • Tandem Mass Spectrometry / methods*

Substances

  • Proteins