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.
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.
Authors: N Leigh Anderson; Malu Polanski; Rembert Pieper; Tina Gatlin; Radhakrishna S Tirumalai; Thomas P Conrads; Timothy D Veenstra; Joshua N Adkins; Joel G Pounds; Richard Fagan; Anna Lobley Journal: Mol Cell Proteomics Date: 2004-01-12 Impact factor: 5.911
Authors: Tyler Stevens; Darwin Conwell; Gregory Zuccaro; Frederick Van Lente; Farah Khandwala; Patrick Hanaway; John J Vargo; John A Dumot Journal: Dig Dis Sci Date: 2004-09 Impact factor: 3.199
Authors: Tyler Stevens; Darwin L Conwell; Gregory Zuccaro; Frederick Van Lente; Farah Khandwala; Edward Purich; John J Vargo; Seymour Fein; John A Dumot; Pat Trolli; Catherine O'Laughlin Journal: Gastrointest Endosc Date: 2004-09 Impact factor: 9.427
Authors: Mads Grønborg; Jakob Bunkenborg; Troels Zakarias Kristiansen; Ole Nørregaard Jensen; Charles J Yeo; Ralph H Hruban; Anirban Maitra; Michael G Goggins; Akhilesh Pandey Journal: J Proteome Res Date: 2004 Sep-Oct Impact factor: 4.466
Authors: Joao A Paulo; Linda S Lee; Bechien Wu; Peter A Banks; Hanno Steen; Darwin L Conwell Journal: J Immunol Methods Date: 2011-05-01 Impact factor: 2.303
Authors: Phil A Hart; Mark Topazian; Massimo Raimondo; Zobeida Cruz-Monserrate; William E Fisher; Gregory B Lesinski; Hanno Steen; Darwin L Conwell Journal: Am J Gastroenterol Date: 2016-08-02 Impact factor: 10.864
Authors: Joao A Paulo; Vivek Kadiyala; Linda S Lee; Peter A Banks; Darwin L Conwell; Hanno Steen Journal: J Proteome Res Date: 2012-02-07 Impact factor: 4.466
Authors: Joao A Paulo; Linda S Lee; Bechien Wu; Peter A Banks; Hanno Steen; Darwin L Conwell Journal: Proteomics Clin Appl Date: 2011-03-01 Impact factor: 3.494
Authors: Darwin L Conwell; Joao A Paulo; Linda S Lee; Peter A Banks; Andrew M Bellizzi; Nisha I Sainani; Vivek Kadiyala; Shadeah Suleiman Journal: J Immunol Methods Date: 2012-06-07 Impact factor: 2.303
Authors: Mohieddin Jafari; Vincent Primo; Gary B Smejkal; Eugene V Moskovets; Winston P Kuo; Alexander R Ivanov Journal: Electrophoresis Date: 2012-08 Impact factor: 3.535
Authors: Joao A Paulo; Linda S Lee; Bechien Wu; Kathryn Repas; Peter A Banks; Darwin L Conwell; Hanno Steen Journal: Proteomics Clin Appl Date: 2010-09 Impact factor: 3.494
Authors: Joao A Paulo; Vivek Kadiyala; Aleksandr Gaun; John F K Sauld; Ali Ghoulidi; Peter A Banks; Hanno Steen; Darwin L Conwell Journal: JOP Date: 2013-03-10