The utility of isotope-coded protein labeling for prioritization of proteins found in ovarian cancer patient urine

Adam Rainczuk, Mark Condina, Matthias Pelzing, Sebastiaan Dolman, Jyothsna Rao, Nicole Fairweather, Tom Jobling, Andrew N. Stephens

Research output: Contribution to journalArticleResearchpeer-review

Abstract

Urine offers a number of attractive features as a sample type for biomarker discovery, including noninvasive sampling, quantity and availability, stability, and a narrow dynamic range. In this study we report the first application of isotope coded protein labeling (ICPL), coupled with in-solution isoelectric fractionation and LC-MALDI-TOF/TOF, to examine and prioritize urinary proteins from ovarian cancer patients. Following the definition of stringent exclusion criteria a total of 579 proteins were identified with 43% providing quantitation data. Protein abundance changes were validated for selected proteins by ESI-Qq-TOF MS, following which Western blot and immunohistochemical analysis by tissue microarray was used to explore the biological relevance of the proteins identified. Several established markers (e.g., HE4, osteopontin) were identified at increased levels in ovarian cancer patient urine, validating the approach used; we also identified a number of potential marker candidates (e.g., phosphatidylethanolamine binding protein 1, cell-adhesion molecule 1) previously unreported in the context of ovarian cancer. We conclude that the ICPL strategy for identification and relative quantitation of urine proteins is an appropriate tool for biomarker discovery studies, and can be applied for the selection of potential biomarker candidates for further characterization.

Original languageEnglish
Pages (from-to)4074-4088
Number of pages15
JournalJournal of Proteome Research
Volume12
Issue number9
DOIs
Publication statusPublished - 6 Sep 2013
Externally publishedYes

Keywords

  • ICPL
  • Isotope-coded protein label
  • ovarian cancer
  • urine

Cite this

Rainczuk, Adam ; Condina, Mark ; Pelzing, Matthias ; Dolman, Sebastiaan ; Rao, Jyothsna ; Fairweather, Nicole ; Jobling, Tom ; Stephens, Andrew N. / The utility of isotope-coded protein labeling for prioritization of proteins found in ovarian cancer patient urine. In: Journal of Proteome Research. 2013 ; Vol. 12, No. 9. pp. 4074-4088.
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The utility of isotope-coded protein labeling for prioritization of proteins found in ovarian cancer patient urine. / Rainczuk, Adam; Condina, Mark; Pelzing, Matthias; Dolman, Sebastiaan; Rao, Jyothsna; Fairweather, Nicole; Jobling, Tom; Stephens, Andrew N.

In: Journal of Proteome Research, Vol. 12, No. 9, 06.09.2013, p. 4074-4088.

Research output: Contribution to journalArticleResearchpeer-review

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