title: Difference gel electrophoresis identifies differentially expressed proteins in endoscopically collected pancreatic fluid.
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.
authors.affiliation: Department of Pathology, Children's Hospital Boston and Harvard Medical School, Boston, MA 02115, USA. Joao.Paulo@childrens.harvard.edu
authors.forename: Joao A
authors.initials: JA
authors.lastname: Paulo
authors.forename: Linda S
authors.initials: LS
authors.lastname: Lee
authors.forename: Peter A
authors.initials: PA
authors.lastname: Banks
authors.forename: Hanno
authors.initials: H
authors.lastname: Steen
authors.forename: Darwin L
authors.initials: DL
authors.lastname: Conwell
mesh_terms: D000328:Adult; D015415:Biomarkers / Q000032:analysis; D002853:Chromatography, Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D016099:Endoscopy, Gastrointestinal; D005260:Female; D006801:Humans; D008297:Male; D008875:Middle Aged; D010186:Pancreatic Function Tests / Q000379:methods*; D010189:Pancreatic Juice / Q000737:chemistry*; D050500:Pancreatitis, Chronic / Q000175:diagnosis / Q000378:metabolism; D010446:Peptide Fragments / Q000032:analysis / Q000378:metabolism; D011506:Proteins / Q000032:analysis* / Q000378:metabolism; D053719:Tandem Mass Spectrometry; D057825:Two-Dimensional Difference Gel Electrophoresis / Q000379:methods*
chemical_list: D015415:Biomarkers; D010446:Peptide Fragments; D011506:Proteins