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Accuracy of two-dimensional electrophoresis for target discovery in human colorectal cancer

The Pharmacogenomics Journal volume 1, pages 142–151 (2001)Cite this article

Abstract

Two-dimensional polyacrylamide gel electrophoresis (2-D PAGE) is increasingly used for target discovery in human disease to complement genomic studies. We have assessed the possibilities and limits of 2-D PAGE applied to human colorectal cancer. Up to 108 epithelial cells were purified from paired normal and pathological biopsies using Ber-EP4 coated magnetic beads, allowing the elimination of cellular and fluid contaminations. The mean coefficient of variation (CVAR) of repeated 2-D PAGE analysis with silver staining was lying between 20 and 28%. However, only 47% (interrun) to 76% (intrarun) of spots could be matched within a triplicate experiment. Interindividual phenotypic variability was high. Intratumoral phenotypic variability was not found to be significant. When method and tumor variability were added, 90% of CVAR were inferior to 48%. Thus, two-fold up- or down-regulation of protein expression reveals biological significance. Serial paired comparison of 923 proteins in 10 patients showed highly reproducible differences between normal and cancer tissues. Under well defined experimental conditions and after the high variability of the technique has been considered, 2-D PAGE parallel analysis of paired colorectal samples allows patient-specific tumor profiling.

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Acknowledgements

We thank I Demalte, JC Sanchez and DF Hochstrasser for significant contributions to this work.

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Authors and Affiliations

  1. Department of Surgery, University of Geneva, Switzerland

    V Ott

  2. Department of Pathology, University of Geneva, Switzerland

    B Borisch

  3. Department of Foundation for Medical Research, University of Geneva, Switzerland

    S Tortola & W Schlegel

  4. Department of Surgery, University of Erlangen, Germany

    K Guenther

  5. Department of Surgery, Academic Hospital Cottbus, Germany

    R Steinert

  6. Department of Surgery, University of Magdeburg, Germany

    M A Reymond

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  1. V Ott
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Ott, V., Guenther, K., Steinert, R. et al. Accuracy of two-dimensional electrophoresis for target discovery in human colorectal cancer. Pharmacogenomics J 1, 142–151 (2001). https://doi.org/10.1038/sj.tpj.6500024

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