Intrinsic indicators for specimen degradation

Jie Li, Catherine Kil, Kelly Considine, Bartosz Smarkucki, Michael C. Stankewich, Brian Balgley, Alexander Vortmeyer

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Variable degrees of molecular degradation occur in human surgical specimens before clinical examination and severely affect analytical results. We therefore initiated an investigation to identify protein markers for tissue degradation assessment. We exposed 4 cell lines and 64 surgical/autopsy specimens to defined periods of time at room temperature before procurement (experimental cold ischemic time (CIT)-dependent tissue degradation model). Using two-dimensional fluorescence difference gel electrophoresis in conjunction with mass spectrometry, we performed comparative proteomic analyses on cells at different CIT exposures and identified proteins with CIT-dependent changes. The results were validated by testing clinical specimens with western blot analysis. We identified 26 proteins that underwent dynamic changes (characterized by continuous quantitative changes, isoelectric changes, and/or proteolytic cleavages) in our degradation model. These changes are strongly associated with the length of CIT. We demonstrate these proteins to represent universal tissue degradation indicators (TDIs) in clinical specimens. We also devised and implemented a unique degradation measure by calculating the quantitative ratio between TDIs' intact forms and their respective degradation-modified products. For the first time, we have identified protein TDIs for quantitative measurement of specimen degradation. Implementing these indicators may yield a potentially transformative platform dedicated to quality control in clinical specimen analyses.

Original languageEnglish (US)
Pages (from-to)242-253
Number of pages12
JournalLaboratory Investigation
Volume93
Issue number2
DOIs
StatePublished - Feb 1 2013
Externally publishedYes

Fingerprint

Cold Ischemia
Proteins
Two-Dimensional Difference Gel Electrophoresis
Quality Control
Proteomics
Proteolysis
Autopsy
Mass Spectrometry
Fluorescence
Western Blotting
Cell Line
Temperature

Keywords

  • alpha-II spectrin
  • degradation
  • indicator
  • proteomics
  • specimen

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Molecular Biology
  • Cell Biology

Cite this

Li, J., Kil, C., Considine, K., Smarkucki, B., Stankewich, M. C., Balgley, B., & Vortmeyer, A. (2013). Intrinsic indicators for specimen degradation. Laboratory Investigation, 93(2), 242-253. https://doi.org/10.1038/labinvest.2012.164

Intrinsic indicators for specimen degradation. / Li, Jie; Kil, Catherine; Considine, Kelly; Smarkucki, Bartosz; Stankewich, Michael C.; Balgley, Brian; Vortmeyer, Alexander.

In: Laboratory Investigation, Vol. 93, No. 2, 01.02.2013, p. 242-253.

Research output: Contribution to journalArticle

Li, J, Kil, C, Considine, K, Smarkucki, B, Stankewich, MC, Balgley, B & Vortmeyer, A 2013, 'Intrinsic indicators for specimen degradation', Laboratory Investigation, vol. 93, no. 2, pp. 242-253. https://doi.org/10.1038/labinvest.2012.164
Li J, Kil C, Considine K, Smarkucki B, Stankewich MC, Balgley B et al. Intrinsic indicators for specimen degradation. Laboratory Investigation. 2013 Feb 1;93(2):242-253. https://doi.org/10.1038/labinvest.2012.164
Li, Jie ; Kil, Catherine ; Considine, Kelly ; Smarkucki, Bartosz ; Stankewich, Michael C. ; Balgley, Brian ; Vortmeyer, Alexander. / Intrinsic indicators for specimen degradation. In: Laboratory Investigation. 2013 ; Vol. 93, No. 2. pp. 242-253.
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