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

Biomarkers / analysis Cold Temperature DNA Damage Endoscopy, Digestive System Freezing Humans Nucleic Acids / analysis Pancreatic Diseases / diagnosis Pancreatic Function Tests Pancreatic Juice / chemistry Predictive Value of Tests Protease Inhibitors / pharmacology Protein Stability Proteins / analysis Proteolysis RNA Stability Ribonucleases / antagonists & inhibitors Secretin / administration & dosage Specimen Handling Time Factors Workflow

Journal

Pancreas

ISSN: 1536-4828

Titre abrégé: Pancreas

Pays: United States

ID NLM: 8608542

Informations de publication

Date de publication:
01 01 2021

Historique:

entrez: 28 12 2020

pubmed: 29 12 2020

medline: 15 12 2021

Statut: ppublish

Résumé

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. 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. 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. 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.

Identifiants

DOI: 10.1097/MPA.0000000000001717 PMID: 33370019 PMC: PMC7883383

pubmed: 33370019

doi: 10.1097/MPA.0000000000001717

pii: 00006676-202101000-00004

pmc: PMC7883383

mid: NIHMS1668851

doi:

Substances chimiques

Biomarkers 0

Nucleic Acids 0

Protease Inhibitors 0

Proteins 0

Secretin 1393-25-5

Ribonucleases EC 3.1.-

Types de publication

Journal Article Research Support, N.I.H., Extramural Research Support, Non-U.S. Gov't

Langues

eng

Sous-ensembles de citation

Pagination

17-28

Subventions

Organisme : NIDDK NIH HHS

ID : U01 DK108326

Pays : United States

Organisme : NIDDK NIH HHS

ID : U01 DK108327

Pays : United States

Informations de copyright

Déclaration de conflit d'intérêts

Z.C.-M., P.A.H., G.B.L., and D.Y. received pilot funds from ChiRhoClin Research Institute, Inc. The rest of the authors declare no conflict of interest.

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