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Molecular & Cellular Proteomics 5:2083-2091, 2006.
© 2006 by The American Society for Biochemistry and Molecular Biology, Inc.

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Research

A Chemical Proteomics Approach for the Identification of Accessible Antigens Expressed in Human Kidney Cancer^*,S

Vincent Castronovo^,, David Waltregny^,, Philippe Kischel, Christoph Roesli^¶, Giuliano Elia^¶, Jascha-N. Rybak^¶,|| and Dario Neri^¶,**

From the Metastasis Research Laboratory, Center of Experimental Cancer Research, University of Liège, 4000 Liège, Belgium and ^¶ Institute of Pharmaceutical Sciences, ETH Zurich, 8093 Zurich, Switzerland

A promising avenue toward the development of more selective anticancer drugs consists in the targeted delivery of bioactive molecules to the tumor environment by means of binding molecules specific to tumor-associated markers. We have used a chemical proteomics approach based on the ex vivo perfusion and biotinylation of accessible structures within surgically resected human kidneys with tumor to gain information about accessible and abundant antigens that are overexpressed in human cancer. This procedure led to the selective labeling with biotin of vascular structures. Biotinylated proteins were purified on streptavidin resin and identified using mass spectrometric methodologies, revealing 637 proteins, 184 of which were only found in tumor specimens and 223 of which were only found in portions of normal kidneys. Immunohistochemical and PCR analysis confirmed that several of the putative cancer antigens identified in this study are indeed preferentially expressed in tumors. In conclusion, we have developed a methodology that allows the identification of accessible biomarkers in human tissues. The tumor-associated antigens identified in this study may be suitable targets for antibody-based anticancer therapies. The experimental approach described here should be applicable to other surgical specimens and to other pathologies as well as to the study of basic physiological and immunological processes.

^** To whom correspondence may be addressed: ETH Zurich, Inst. of Pharmaceutical Sciences, HCI G 396, Wolfgang-Pauli-Strasse 10, 8093 Zurich, Switzerland. Tel.: 41-44-63-37401; Fax: 41-44-63-31358; E-mail: neri{at}pharma.ethz.ch

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