Use of cancer-specific yeast-secreted in vivo biotinylated recombinant antibodies for serum biomarker discovery
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BioMed Central
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Research
Use of cancer-specific yeast-secreted in vivo biotinylated recombinant antibodies for serum biomarker discovery Nathalie Scholler*1, Jennifer A Gross2, Barbara Garvik2, Lance Wells3, Yan Liu2, Christian M Loch2, Arturo B Ramirez2, Martin W McIntosh2, Paul D Lampe2 and Nicole Urban2 Address: 1Center for Research on Early Detection and Cure of Ovarian Cancer, School of Medicine, University of Pennsylvania, Philadelphia, Pennsylvania 19104, USA, 2Molecular Diagnostics Program, Public Health Sciences, Fred Hutchinson Cancer Research Center, Seattle, Washington 98109, USA and 3Complex Carbohydrate Research Center, University of Georgia, Athens, Georgia 30602, USA Email: Nathalie Scholler* - [email protected]; Jennifer A Gross - [email protected]; Barbara Garvik - [email protected]; Lance Wells - [email protected]; Yan Liu - [email protected]; Christian M Loch - [email protected]; Arturo B Ramirez - [email protected]; Martin W McIntosh - [email protected]; Paul D Lampe - [email protected]; Nicole Urban - [email protected] * Corresponding author
Published: 24 July 2008 Journal of Translational Medicine 2008, 6:41
doi:10.1186/1479-5876-6-41
Received: 7 March 2008 Accepted: 24 July 2008
This article is available from: http://www.translational-medicine.com/content/6/1/41 © 2008 Scholler et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
Abstract Background: Strategies to discover circulating protein markers of ovarian cancer are urgently needed. We developed a novel technology that permits us to isolate recombinant antibodies directed against the potential serum biomarkers, to facilitate the further development of affinity reagents necessary to construct diagnostic tests. Methods: This study presents a novel discovery approach based on serum immunoprecipitation with cancer-specific in vivo biotinylated recombinant antibodies (biobodies) derived from differentially selected yeast-display scFv, and analysis of the eluted serum proteins by electrophoresis and/or mass spectrometry. Results: Using this strategy we identified catabolic fragments of complement factors, EMILIN2, Von Willebrand factor and phosphatidylethanolamine-binding protein 1 (PEBP1 or RKIP) in patient sera. To our knowledge, this is the first report of a soluble form of PEBP1 in human. Independent evidence for ovarian cancer-specific expression of PEBP1 in patient sera was found by ELISA assays and antibody arrays with anti-PEBP1 antibodies. PEBP1 was detected in 29 out of 30 ascites samples and discriminated ovarian cancer sera from controls (p = 0.02). Finally, we confirmed by western blots the presence of a 21–23 kDa fragment corresponding to the expected size of PEBP1 but we also showed additional bands of 38 kDa and 50–52 kDa in various tissues and
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