A novel technique to specifically analyze the secretome of cells and tissues

Hannes Zwickl, Elisabeth Traxler, Stefan Staettner, Wolfram Parzefall, Bettina Grasl-Kraupp, Josef Karner, Rolf Schulte-Hermann, Christopher Gerner

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70 Citations (Scopus)

Abstract

The secretome of cells and tissues may reflect a broad variety of pathological conditions and thus represents a rich source of biomarkers. The identity of secreted proteins, usually isolated from cell supernatants or body fluids, is hardly accessible by direct proteome analysis, because these proteins are often masked by high amounts of proteins actually not secreted by the investigated cells. Here, we present a novel method for the specific detection of proteins secreted by human tissue specimen as well as cultured cells and chose liver as a model. The method is based on the metabolic labelling of proteins synthesized during a limited incubation period. Then, the cell supernatant is filtered, precipitated, and subjected to two-dimensional gel electrophoresis. Whereas fluorography detected a large number of proteins derived from residual plasma and dead cells, the autoradiographs selectively displayed genuinely secreted proteins. We demonstrate the feasibility of this approach by means of the secretomes of the hepatocellular carcinoma-derived cell line HepG2 and human liver slices. The selective identification of cell- and tissue-specific protein secretion profiles may help to identify novel sets of biomarkers for wide clinical applications.

Original languageEnglish
Pages (from-to)2779-2785
Number of pages7
JournalELECTROPHORESIS
Volume26
Issue number14
DOIs
Publication statusPublished - Jul 1 2005

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Keywords

  • Biomarker discovery
  • Liver
  • Metabolic labelling
  • Proteomics
  • Secretome

ASJC Scopus subject areas

  • Analytical Chemistry
  • Biochemistry
  • Clinical Biochemistry

Cite this

Zwickl, H., Traxler, E., Staettner, S., Parzefall, W., Grasl-Kraupp, B., Karner, J., Schulte-Hermann, R., & Gerner, C. (2005). A novel technique to specifically analyze the secretome of cells and tissues. ELECTROPHORESIS, 26(14), 2779-2785. https://doi.org/10.1002/elps.200410387