Identification of Pancreas-Specific Proteins in Endoscopically (Endoscopic Pancreatic Function Test) Collected Pancreatic Fluid with Liquid Chromatography- Tandem Mass Spectrometry

Abstract

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. 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. 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. 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.