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Séminaire présenté par Yannis François, LDSM2, Strasbourg

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Sheathless Capillary Electrophoresis – Mass Spectrometry coupling as a flexible platform : From nanoESI infusion device for noncovalent complexes characterization to peptide mapping of monoclonal antibodies

Par : Yannis François, LDSM2, UMR7177, Strasbourg
Date : vendredi 24 mai 2013 à 11h
Lieu : IPHC, Amphithéâtre Grünewald, Bâtiment 25

Résumé :

R. Gahoual1, J.M. Busnel2, L. Kuhn3, P. Hammann3, Y.N. François1, E. Leize-Wagner1

1 Laboratoire de Dynamique et Structure Moléculaire par Spectrométrie de Masse (LDSM2), CNRS – UMR7177, University of Strasbourg, Strasbourg
2 Beckman Coulter Inc., Brea, Californie, Etats-Unis
3 Plateforme protéomique, Université de Strasbourg, Institut de Biologie Moléculaire et Cellulaire, Strasbourg, France

Full protein characterization, as now requested by regulation agencies, is still an actual challenge for analytical sciences. Capillary Electrophoresis (CE) coupled with Mass Spectrometry (MS) is a technique highly suitable for the separation and detection of intact proteins and peptide mixtures. Since 1988, several approaches for coupling CE to MS have been described. However, due to its miniaturized format ; CE suffers from a lower loading capacity and a difficulty to maintain the electrical field.

A novel sheathless CE-ESI-MS platform, referred to as CESI-MS, allowing hyphenation of CE to electrospray ionization MS (ESI-MS), has been developed by Beckman Coulter. This CESI prototype uses a bare fused-silica capillary whose outlet has been etched using hydrofluoric acid making it porous to the electrical transport of small ions without permitting any other significant matter transfer through the pores. The porous tip provides electrical contact by means of a second capillary. Here, we have evaluated the suitability of CESI-MS platform as the nanoelectrospray emitter to analyze with high sensitivity intact proteins and non-covalent complexes. On the other hand, CESI-MS system has been used to develop a CE-MS/MS method allowing a fast and precise characterization of a monoclonal Antibody (mAb) digest.

Personne à contacter : Quentin RAFFY