Difference gel electrophoresis identifies differentially expressed proteins in endoscopically collected pancreatic fluid

Joao A. Paulo, Linda S. Lee, Peter A. Banks, Hanno Steen, Darwin L. Conwell

Research output: Contribution to journalArticlepeer-review

7 Scopus citations

Abstract

Alterations in the pancreatic fluid proteome of individuals with chronic pancreatitis (CP) may offer insights into the development and progression of the disease. The endoscopic pancreatic function test (ePFT) can safely collect large volumes of pancreatic fluid that are potentially amenable to proteomic analyses using difference gel electrophoresis (DIGE) coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS). Pancreatic fluid was collected endoscopically using the ePFT method following secretin stimulation from three individuals with severe CP and three chronic abdominal pain (CAP) controls. The fluid was processed to minimize protein degradation and the protein profiles of each cohort, as determined by DIGE and LC-MS/MS, were compared. This DIGE-LC-MS/MS analysis reveals proteins that are differentially expressed in CP compared with CAP controls. Proteins with higher abundance in pancreatic fluid from CP individuals include: actin, desmoplankin, α-1-antitrypsin, SNC73, and serotransferrin. Those of relatively lower abundance include carboxypeptidase B, lipase, α-1-antichymotrypsin, α-2-macroglobulin, actin-related protein (Arp2/3) subunit 4, glyceraldehyde-3-phosphate dehydrogenase, and protein disulfide isomerase. Endoscopic collection (ePFT) in tandem with DIGE-LC-MS/MS is a suitable approach for pancreatic fluid proteome analysis; however, further optimization of our protocol, as outlined herein, may improve proteome coverage in future analyses.

Original languageEnglish
Pages (from-to)1939-1951
Number of pages13
JournalElectrophoresis
Volume32
Issue number15
DOIs
StatePublished - Aug 2011

Keywords

  • Biomarkers
  • Pancreas
  • Pancreatic function test
  • Pancreatic juice

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Fingerprint

Dive into the research topics of 'Difference gel electrophoresis identifies differentially expressed proteins in endoscopically collected pancreatic fluid'. Together they form a unique fingerprint.

Cite this