New methodologies in native mass spectrometry and ion mobility for the structural characterization of proteins of therapeutic interest and multiprotein complexes
Description
This PhD work focuses on developments in native mass spectrometry and ion mobility methods for the structural characterization of therapeutic proteins and multiprotein complexes. First, careful optimizations of sample preparation and analytical conditions allowed the characterization of membrane proteins, which are hydrophobic proteins difficult to analyze by MS approaches in detergent environment. Then, a new ion mobility-based activation approach called Collision Induced Unfolding has been set up and evaluated. CIU allowed extensive and original conformational characterization of several therapeutic monoclonal antibody formats. Finally, native MS and ion mobility techniques were used for the characterization of heterogeneous multiprotein complexes depicting their benefit when combined to other biophysical techniques for the structural characterization of multiprotein complexes. (author)
Abstract (French)
Ce travail de these repose sur le developpement de methodes en spectrometrie de masse native et mobilite ionique pour la caracterisation structurale de proteines d'interet therapeutique et de complexes multiproteiques. L'optimisation fine et consequente de la preparation d'echantillon et des conditions analytiques ont permis la caracterisation de proteines membranaires solubilisees en milieu detergent, proteines hydrophobes habituellement refractaires a l'analyse par MS. D'autre part, une nouvelle approche de mobilite ionique appelee Collision Induced Unfolding a ete evaluee et mise en place au laboratoire. Elle a permis une caracterisation conformationnelle approfondie et originale de plusieurs formats d'anticorps monoclonaux therapeutiques. Enfin, les techniques de MS native et de mobilite ionique ont ete utilisees pour caracteriser des complexes multiproteiques d'heterogeneite variable mettant ainsi en lumiere leurs avantages et les progres realises dans le domaine de la MS structurale. (auteur)
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Additional details
Additional titles
- Original title (French)
- Nouvelles methodologies en spectrometrie de masse native et mobilite ionique pour la caracterisation structurale de proteines d'interet therapeutique et de complexes multiproteiques
Publishing Information
- Imprint Pagination
- 278 p.
- Report number
- FRNC-TH--12628
INIS
- Country of Publication
- France
- Country of Input or Organization
- France
- INIS RN
- 53029655
- Subject category
- S37: INORGANIC, ORGANIC, PHYSICAL AND ANALYTICAL CHEMISTRY; S46: INSTRUMENTATION RELATED TO NUCLEAR SCIENCE AND TECHNOLOGY; S60: APPLIED LIFE SCIENCES;
- Resource subtype / Literary indicator
- Thesis
- Descriptors DEI
- CHARGE STATES; COMPLEXES; ELECTRIC POTENTIAL; ION MOBILITY; LIGANDS; MASS SPECTROSCOPY; MEMBRANE PROTEINS; MONOCLONAL ANTIBODIES; PH VALUE; QUADRUPOLES; TIME-OF-FLIGHT MASS SPECTROMETERS
- Descriptors DEC
- ANTIBODIES; DYNAMIC MASS SPECTROMETERS; MASS SPECTROMETERS; MEASURING INSTRUMENTS; MOBILITY; MULTIPOLES; ORGANIC COMPOUNDS; PARTICLE MOBILITY; PROTEINS; SPECTROMETERS; SPECTROSCOPY; TIME-OF-FLIGHT SPECTROMETERS
Optional Information
- Notes
- 393 refs.; Available from the INIS Liaison Officer for France, see the INIS website for current contact and E-mail addresses