The impact of pre-analytical conditions on the serum proteome: heat-stabilization versus nitrogen storage

Arch Physiol Biochem. 2013 Jul;119(3):100-7. doi: 10.3109/13813455.2013.806556. Epub 2013 Jul 4.

Abstract

Context: Biological material reflecting the in vivo composition of markers provides a high potential for biomarker discovery.

Objective: We compared the serum proteome following heat- and nitrogen-preservation, with and without subsequent storage at room temperature.

Materials and methods: Serum samples were collected, treated and analysed by two-dimensional gel electrophoresis. Protein spots were identified and confirmed by two mass spectrometry approaches (MALDI & ESI) and subjected to Ingenuity Pathway Analysis.

Results: We revealed 24 differentially expressed proteins (p ≤ 0.05) between nitrogen and heat preservation, and 87 between nitrogen and heat preservation with subsequent storage for 120 h at room-temperature. Mass spectrometry identified 25 polypeptides. Pathway analysis resulted in networks maintaining Cellular Assembly and Organization, Movement and Maintenance.

Conclusion: Heat-stabilization does not substantially change the short-term proteome composition of serum compared with nitrogen treatment. However, heat-stabilization alone seems insufficient for long-term sample preservation for serum samples. We identified transthyretin and apolipoprotein A-IV as sample quality markers.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / blood
  • Blood Proteins / analysis*
  • Colonic Neoplasms / blood*
  • Colonic Neoplasms / diagnosis
  • Cryopreservation*
  • Electrophoresis, Gel, Two-Dimensional
  • Hot Temperature
  • Humans
  • Nitrogen
  • Principal Component Analysis
  • Protein Stability
  • Proteomics / standards*
  • Reproducibility of Results
  • Spectrometry, Mass, Electrospray Ionization
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • Biomarkers
  • Blood Proteins
  • Nitrogen