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Comprehensive Proteomic Analysis of Human Pancreatic Juice

Mads Grønborg,^†^‡ Jakob Bunkenborg,^†^‡ Troels Zakarias Kristiansen,^†^‡ Ole Nørregaard Jensen,^‡ Charles J. Yeo,^§ Ralph H. Hruban, Anirban Maitra, Michael G. Goggins, and Akhilesh Pandey*^†

McKusick-Nathans Institute of Genetic Medicine and Department of Biological Chemistry, Johns Hopkins University, Baltimore, Maryland 21205, Department of Biochemistry and Molecular Biology, University of Southern Denmark, Campusvej 55, Odense M, Denmark, Department of Surgery, Johns Hopkins University, Baltimore, Maryland 21287, and Department of Pathology, Johns Hopkins University, Baltimore, Maryland 21287

Journal of Proteome Research, 2004, 3 (5), pp 1042–1055

DOI: 10.1021/pr0499085

Publication Date (Web): September 17, 2004

†

McKusick-Nathans Institute of Genetic Medicine and Department of Biological Chemistry, Johns Hopkins University.

‡

Department of Biochemistry and Molecular Biology, University of Southern Denmark.

Department of Surgery, Johns Hopkins University.

Department of Pathology, Johns Hopkins University.

To whom correspondence should be addressed. E-mail: pandey@ jhmi.edu.

Abstract

Proteomic technologies provide an excellent means for analysis of body fluids for cataloging protein constituents and identifying biomarkers for early detection of cancers. The biomarkers currently available for pancreatic cancer, such as CA19-9, lack adequate sensitivity and specificity contributing to late diagnosis of this deadly disease. In this study, we carried out a comprehensive characterization of the “pancreatic juice proteome” in patients with pancreatic adenocarcinoma. Pancreatic juice was first fractionated by 1-dimensional gel electrophoresis and subsequently analyzed by liquid chromatography tandem mass spectrometry (LC−MS/MS). A total of 170 unique proteins were identified including known pancreatic cancer tumor markers (e.g., CEA, MUC1) and proteins overexpressed in pancreatic cancers (e.g., hepatocarcinoma−intestine−pancreas/pancreatitis-associated protein (HIP/PAP) and lipocalin 2). In addition, we identified a number of proteins that have not been previously described in pancreatic juice (e.g., tumor rejection antigen (pg96) and azurocidin). Interestingly, a novel protein that is 85% identical to HIP/PAP was identified, which we have designated as PAP-2. The proteins identified in this study could be directly assessed for their potential as biomarkers for pancreatic cancer by quantitative proteomics methods or immunoassays.

Keywords: liquid chromatography tandem mass spectrometry (LC−MS/MS) • functional annotation • body fluid

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