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<dc:title xml:lang="fr">Développement de méthodes de préparation d'échantillons et d'analyse LC-MS/MS pour l'étude de protéomes de cellules uniques</dc:title>
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<dc:subject xml:lang="fr">Acquisiton indépendante des données-accumulaton parallèle et fragmentaton en série (DIA-PASEF)</dc:subject>
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<dcterms:abstract xml:lang="fr">La spectrométrie de masse est une méthode de choix pour analyser le protéome, permettant d’identifier et de quantifier à haute résolution les protéines d’un mélange complexe. Les avancées récentes rendent désormais envisageable l’étude du protéome à l’échelle unicellulaire (scP), offrant une compréhension fine de l’hétérogénéité cellulaire, jusqu’alors masquée par l’approche en bulk.Toutefois, son application reste limitée en raison des défis liés aux quantités infimes de protéines disponibles par cellule unique (80-400pg). Dans ce contexte, cette thèse s’est consacrée au développement de méthodes de préparation d’échantillon et d’analyse nano LC-MS/MS dédiée à la scP. Ces optimisations ont ensuite été appliquées à des modèles biologiques, notamment dans le cadre de l’étude de l’hétérogénéité cellulaire lors d’une infection virale et le sous-typage de cellules immunitaires.</dcterms:abstract>
<dcterms:abstract xml:lang="en">Mass spectrometry is a method of choice for analyzing the proteome, enabling high-resoluton identfcaton and quantfcaton of proteins in a complex mixture. Recent advances now make it possible to study the proteome at the single-cell level (scP), providing a detailed understanding of cellular heterogeneity, which was previously masked by the bulk approach. However, its applicaton remains limited due to challenges related to the tny amounts of protein available per single cell(80-400 pg). In this context, this thesis work focused on the development of sample preparaton and nano LC-MS/MS analysis methods dedicated to scP. These optmizatons were then applied to biological models, partcularly in the study of cellular heterogeneity during viral infecton and the subtyping of immune cells.</dcterms:abstract>
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