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A more recent version of this article appeared on August 1, 2005. Originally published In Press as doi:10.1074/mcp.M400175-MCP200 on May 18, 2005.
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Submitted on November 10, 2004
Revised on May 13, 2005
Accepted on May 17, 2005

An immunoproteomic approach for identification of clinical biomarkers for monitoring disease: Application to cystic fibrosis

Susanne K. Pedersen, Andrew J. Sloane, Sindhu S. Prasad, Lucille T. Sebastian, Robyn A. Lindner, Michael Hsu, Michael Robinson, Peter T. Bye, Ron P. Weinberger, and Jenny L. Harry

Discovery, Proteome Systems, Sydney, NSW 2113

Corresponding Author: sanne.pedersen{at}proteomesystems.com

Circulating antibodies can be used to probe protein arrays of body fluids, prepared by two-dimensional gel electrophoresis (2-DE), for antigenic biomarker detection. However, detected proteins, particularly low abundance antigens, often remain unidentifiable due to proteome complexity and limiting sample amounts. Using a novel enrichment approach exploiting patient antibodies for isolation of antigenic biomarkers, we demonstrate how immunoproteomic strategies can accelerate biomarker discovery. Application of this approach as a means of identifying biomarkers was demonstrated for cystic fibrosis (CF) lung disease by isolation and identification of inflammatory-associated autoantigens, including myeloperoxidase and calgranulin B from sputum of subjects with CF. The approach was also exploited for isolation of proteins expressed by the Pseudomonas aeruginosa strain PA01. Capture of PA01 antigens using circulating antibodies from CF subjects implicated in vivo expression of Pseudomonas proteins. All CF subjects screened, but not controls, were immunoreactive against immuno-captured Pseudomonas proteins, representing stress- (groES and ferric iron-binding protein Hit A), immunosuppressive- (thioredoxin) and alginate synthetase pathway- (nucleoside diphosphate kinase) proteins, implicating their clinical relevance as biomarkers of infection.


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