Proteomic Analyses of Pancreatic Cyst Fluids

From the Divisions of *Basic Science, daggerMedical Science, and double daggerPopulation Science, Fox Chase Cancer Center, Philadelphia, PA.
Pancreas (Impact Factor: 2.96). 02/2009; 38(2):e33-42. DOI: 10.1097/MPA.0b013e318193a08f
Source: PubMed

ABSTRACT There are currently no diagnostic indicators that are consistently reliable, obtainable, and conclusive for diagnosing and risk-stratifying pancreatic cysts. Proteomic analyses were performed to explore pancreatic cyst fluids to yield effective diagnostic biomarkers.
We have prospectively recruited 20 research participants and prepared their pancreatic cyst fluids specifically for proteomic analyses. Proteomic approaches applied were as follows: (1) matrix-assisted laser-desorption-ionization time-of-flight mass spectrometry peptidomics with LC/MS/MS (HPLC-tandem mass spectrometry) protein identification; (2) 2-dimensional gel electrophoresis; (3) GeLC/MS/MS (tryptic digestion of proteins fractionated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by LC/MS/MS).
Sequencing of more than 350 free peptides showed that exopeptidase activities rendered peptidomics of cyst fluids unreliable; protein nicking by proteases in the cyst fluids produced hundreds of protein spots from the major proteins, making 2-dimensional gel proteomics unmanageable; GeLC/MS/MS revealed a panel of potential biomarker proteins that correlated with carcinoembryonic antigen (CEA).
Two homologs of amylase, solubilized molecules of 4 mucins, 4 solubilized CEA-related cell adhesion molecules (CEACAMs), and 4 S100 homologs may be candidate biomarkers to facilitate future pancreatic cyst diagnosis and risk-stratification. This approach required less than 40 microL of cyst fluid per sample, offering the possibility to analyze cysts smaller than 1 cm in diameter.

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    Gastroenterology Research and Practice 01/2012; 2012(2):147465. DOI:10.1155/2012/147465 · 1.75 Impact Factor
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    • "Fluid was analyzed using MALDI-TOF mass spectrometry with LC/MS/MS protein identification, 2D gel electrophoresis, or GeLC/MS/MS (tryptic digestion of proteins fractionated by SDS-PAGE and identified by LC/MS/MS). The first two techniques proved to be unsatisfactory, presumably from endogenous peptidases which splintered native proteins in numerous locations [81]. Mass spectrometry yielded homologs within three families of proteins associated with pancreatic cancer, including mucins, CEACAMs, and S100s. "
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