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Séminaire Chimie ED459

New mass spectrometry approaches for detailed characterization of proteins and their biological complexes

Dr. Emmanuelle Leize-Wagner, D.R. CNRS (LSMIS Labo de Spectrométrie de Masse des Interactions et des Systèmes, UMR 7140 Chimie de la Matière Complexe, CNRS, Université de Strasbourg)

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Le Jeudi 24 novembre 2016 à 13h45
UM FdS, Salle de Cours SC-16.01

LSIMS’s research activities are focussed on the development of mass spectrometry (MS) for the characterization of proteins related to :

A. The development of MS to improve the structural and dynamic study of their protein complexes with the determination of their stoichiometry, their stability constants, their interaction areas (“cross-linking”), and that working on increasingly heavy systems (several millions Dalton) and on increasingly small sample amounts (a few picomoles). Today native MS is primarily a tool for determining the stoichiometry of the complexes and the nature of the partners involved in the complexes. It is rarely used for following the complexes formation (kinetic study) and for determining their stability constants. So our goal is through the study of different systems to determine the conditions under which MS can obtain such information by demonstrating in particular that the mass spectrum gives the qualitative and quantitative image of the species present in solution (including comparison of the results obtained with other techniques). The deep characterization of these complexes involves also defining their interacting areas. Understanding the way how proteins interact with each other to form transient or stable protein complexes is a key aspect in structural biology. In this project, we combine chemical cross-linking with mass spectrometry (CX-MS) to map the protein-protein interaction network of the complexes.

B. The development of MS to improve the characterization of intact proteins. In the case of therapeutic proteins, such as monoclonal antibodies (mAbs), complete protein characterization is now requested by regulation agencies. In this context, we have developed over the last few years new MS strategies primarily based on the novel sheathless coupling of capillary electrophoresis with electrospray mass spectrometry (CESI-MS). mAbs have become one of the most rapidly growing classes of therapeutic proteins in the treatment of human diseases. mAbs are highly heterogeneous proteins (glycosylations, post-translationnal modifications), thereby requiring a large set of sophisticated analytical technologies for their complete characterization. For the first time, CESI-MS enables the access, in a single injection and with less than one picomole of protein, to the complete structure (including all the glycosylations and modifications) of mAbs and related products (biosimilars, Antibody-Drug Conjugates ADCs).

Contact local IBMM : Prof. Christine Enjalbal (DAPP)

Voir en ligne : LSMIS website


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