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Développement et validation de méthodes de protéomique innovantes pour des applications de biochimie clinique

Abstract : In recent years, mass spectrometry has been considered in the analytical chemistry field as the reference method. "Proteomics", a concept that emerged in the 2000s, consists in the identification and/or quantification of peptides and proteins in different type of samples (cells, tissues, or biological samples), in various conditions. A more specific field of this concept, the "clinical proteomics" specifically concerns the study of proteome for the research on one hand, diagnostic markers, prognostic and therapeutic follow-up of human pathologies and, on the other hand, of pathophysiological actors that can serve as a therapeutic target.Currently, the technology of choice used for the analysis of proteins in clinical biochemistry is the ELISA (Enzyme-linked immunosorbent assay) which has major drawbacks: not a multiplex analysis, high variability, no standardization, and the inability to distinguish proteoforms of one protein. The targeted proteomics by Liquid Chromatography/Multiple Reaction Monitoring (LC-MRM) allows to surpass these disadvantages because is highly multiplexable (> 200 proteins/analysis), robust, compatible to the use of protein/peptide standards, and is able to distinguish a wide variety of post translational modifications.In this thesis project, evaluation and validation of the targeted mass spectrometry (LC-MRM) will be perform for the quantification of proteins with clinical interest. In this context, we will present three clinical proteomics developments: apolipoprotein E phenotyping, considered as the best risk factor for Alzheimer's disease; Monoclonal therapeutic antibody (Bevacizumab) quantification in patient serum; and absolute quantification of hepcidin-25 in the context of diseases related to iron metabolism.
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Jerome Vialaret. Développement et validation de méthodes de protéomique innovantes pour des applications de biochimie clinique. Sciences agricoles. Université Montpellier, 2019. Français. ⟨NNT : 2019MONTT038⟩. ⟨tel-02479366⟩

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