Pre-fractionation of Noncirculating Biological Fluids to Improve Discovery of Clinically Relevant Protein Biomarkers
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Nowadays, significant difficulties remain in the diagnosis and/or prognosis of many diseases, leading to an unsatisfactory patient management and a counterproductive increase in time and costs. It is therefore crucial to bridge the gap between basic and applied research by complying with clinical requirements, notably from the design stage of the experimental workflow. In this chapter we provide key suggestions for selecting appropriate biological samples and reducing pre-analytical and analytical variabilities to improve the discovery of clinically relevant protein biomarkers.
Key wordsDifferential centrifugation Extracellular Vesicles Exosomes Microvesicles Body fluids Differential diagnosis Translational medicine
The author sincerely thanks all the coworkers who, over the years, have substantially contributed in the acquisition of the know-how required for the development of the present protocols, notably: Dr. Yohann Couté (Exploring the Dynamics of Proteomes laboratory at the CEA, Grenoble, France) for the methodological advices about each phase of protein identification and quantitation; Dr. Valeria Severino (Digestive Cancers Biomarkers Group of the Medicine Faculty at the Geneva University, Geneva, Switzerland) for the improvement of protein sub-fractionation protocols; The Proteomic Core Facility (Medicine Faculty at the Geneva University, Geneva, Switzerland) for the adaptation of peptide cleanup methods; Dr. Pierre Lescuyer, (Department of Genetic, Laboratory and Pathology Medicine at the Geneva University Hospitals, Geneva, Switzerland) for crucial knowledge in clinical laboratory requirements; Dr. Jean-Marc Dumonceau (Gedyt Center, Argentina, Buenos Aires) for essential expertise in sample inclusion and collection; Prof. Jean-Louis Frossard (Gastroenterology and Hepatology Service at the Geneva University Hospitals, Geneva, Switzerland) for strong competences in digestive pathologies, constructive criticisms, and continuous help and support; all the trainee students involved in the analyses that allowed assembling and validating the protocols.
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