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J Physiol Volume 563, Number 1, 1-9, February 15, 2005 DOI: 10.1113/jphysiol.2004.080341
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Topical Reviews

Broad-based proteomic strategies: a practical guide to proteomics and functional screening

David R. M Graham1,2, Steven T Elliott1 and Jennifer E Van Eyk1,3,4

Departments of
1 Medicine, Division of Cardiology
2 Pharmacology and Molecular Sciences
3 Biomedical Engineering
4 Biological Chemistry, Johns Hopkins University, Baltimore, MD, USA

Proteomics, the study of the proteome (the collection of all the proteins expressed from the genome in all isoforms, polymorphisms and post-translational modifications), is a rapidly developing field in which there are numerous new and often expensive technologies, making it imperative to use the most appropriate technology for the biological system and hypothesis being addressed. This review provides some guidelines on approaching a broad-based proteomics project, including strategies on refining hypotheses, choosing models and proteomic approaches with an emphasis on aspects of sample complexity (including abundance and protein characteristics), and separation technologies and their respective strengths and weaknesses. Finally, issues related to quantification, mass spectrometry and informatics strategies are discussed. The goal of this review is therefore twofold: the first section provides a brief outline of proteomic technologies, specifically with respect to their applications to broad-based proteomic approaches, and the second part provides more details about the application of these technologies in typical scenarios dealing with physiological and pathological processes. Proteomics at its best is the integration of carefully planned research and complementary techniques with the advantages of powerful discovery technologies that has the potential to make substantial contributions to the understanding of disease and disease processes.

(Received 1 December 2004; accepted after revision 15 December 2004; first published online 16 December 2004)
Corresponding author J. E. Van Eyk: 602 Mason F. Lord Bldg, Center Tower, 5200 Eastern Ave, Johns Hopkins University, Baltimore, MD 21224, USA. Email: jvaneyk1{at}jhmi.edu




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