Delayed Processing of Secretin-Induced Pancreas Fluid Influences the Quality and Integrity of Proteins and Nucleic Acids

Zobeida Cruz-Monserrate, Kristyn Gumpper, Sabrina Kaul, Niharika Badi, Samantha Terhorst, Kelly Dubay, Gregory B. Lesinski, William Fisher, Amy McElhany, Luis F. Lara, Somashekar Krishna, Thomas Mace, Natalia Higuita-Castro, Lilibeth Ortega-Pineda, Michael A. Freitas, Alice Hinton, Dhiraj Yadav, Phil A. Hart, Stephen J. Pandol, Saima AhmedBenoit Fatou, Hanno Steen, Darwin L. Conwell

Research output: Contribution to journalArticlepeer-review

4 Scopus citations


Objectives Endoscopic pancreatic function tests are used to diagnose pancreatic diseases and are a viable source for the discovery of biomarkers to better characterize pancreatic disorders. However, pancreatic fluid (PF) contains active enzymes that degrade biomolecules. Therefore, we tested how preservation methods and time to storage influence the integrity and quality of proteins and nucleic acids. Methods We obtained PF from 9 subjects who underwent an endoscopic pancreatic function test. Samples were snap frozen at the time of collection; after 1, 2, and 4 hours on ice; or after storage overnight at 4°C with or without RNase or protease inhibitors (PIs). Electrophoresis and mass spectrometry analysis determined protein abundance and quality, whereas nucleic acid integrity values determined DNA and RNA degradation. Results Protein degradation increased after 4 hours on ice and DNA degradation after 2 hours on ice. Adding PIs delayed degradation. RNA was significantly degraded under all conditions compared with the snap frozen samples. Isolated RNA from PF-derived exosomes exhibited similar poor quality as RNA isolated from matched PF samples. Conclusions Adding PIs immediately after collecting PF and processing the fluid within 4 hours of collection maintains the protein and nucleic acid integrity for use in downstream molecular analyses.

Original languageEnglish
Pages (from-to)17-28
Number of pages12
Issue number1
StatePublished - 2021

Bibliographical note

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  • biomarkers
  • pancreatic cancer
  • pancreatic fluid
  • pancreatitis
  • sample degradation

ASJC Scopus subject areas

  • Internal Medicine
  • Endocrinology, Diabetes and Metabolism
  • Hepatology
  • Endocrinology


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