title: Toward a suitable structural analysis of gene delivery carrier based on polycationic carbohydrates by electron transfer dissociation tandem mass spectrometry.
abstract: Polycationic carbohydrates represent an attractive class of biomolecules for several applications and particularly as non viral gene delivery vectors. In this case, the establishment of structure-biological activity relationship requires sensitive and accurate characterization tools to both control and achieve fine structural deciphering. Electrospray-tandem mass spectrometry (ESI-MS/MS) appears as a suitable approach to address these questions. In the study herein, we have investigated the usefulness of electron transfer dissociation (ETD) to get structural data about five polycationic carbohydrates demonstrated as promising gene delivery agents. A particular attention was paid to determine the influence of charge states as well as both fluoranthene reaction time and supplementary activation (SA) on production of charge reduced species, fragmentation yield, varying from 2 to 62%, as well as to obtain the most higher both diversity and intensity of fragments, according to charge states and targeted compounds. ETD fragmentation appeared to be mainly directed toward pending group rather than carbohydrate cyclic scaffold leading to a partial sequencing for building blocks when amino groups are close to carbohydrate core, but allowing to complete structural deciphering of some of them, such as those including dithioureidocysteaminyl group which was not possible with CID only. Such findings clearly highlight the potential to help the rational choice of the suitable analytical conditions, according to the nature of the gene delivery molecules exhibiting polycationic features. Moreover, our ETD-MS/MS approach open the way to a fine sequencing/identification of grafted groups carried on various sets of oligo-/polysaccharides in various fields such as glycobiology or nanomaterials, even with unknown or questionable extraction, synthesis or modification steps.
authors.affiliation: Université d'Evry-Val-d'Essonne, Laboratoire Analyse et Modélisation pour la Biologie et l'Environnement, CNRS UMR 8587, Bâtiment Maupertuis, Bld F. Mitterrand, F-91025 Evry, France. Electronic address: cedric.przybylski@upmc.fr.
authors.forename: Cédric
authors.initials: C
authors.lastname: Przybylski
authors.affiliation: Instituto de Investigaciones Químicas (IIQ), CSIC-Universidad de Sevilla, Américo Vespucio 49, Isla de la Cartuja, E-41092 Sevilla, Spain.
authors.forename: Juan M
authors.initials: JM
authors.lastname: Benito
authors.affiliation: Université de Picardie Jules Verne, Laboratoire de Glycochimie, des Antimicrobiens et des Agroressources, CNRS UMR 7378, 80039 Amiens, France.
authors.forename: Véronique
authors.initials: V
authors.lastname: Bonnet
authors.affiliation: Departamento de Química Orgánica, Facultad de Química, Universidad de Sevilla, E-41012 Sevilla, Spain.
authors.forename: Carmen Ortiz
authors.initials: CO
authors.lastname: Mellet
authors.affiliation: Instituto de Investigaciones Químicas (IIQ), CSIC-Universidad de Sevilla, Américo Vespucio 49, Isla de la Cartuja, E-41092 Sevilla, Spain.
authors.forename: José M
authors.initials: JM
authors.lastname: García Fernández
mesh_terms: D002236:Carbohydrate Conformation; D002241:Carbohydrates / Q000737:chemistry*; D004583:Electrons; D018014:Gene Transfer Techniques; D011073:Polyamines / Q000737:chemistry*; D000071228:Polyelectrolytes; D021241:Spectrometry, Mass, Electrospray Ionization / Q000379:methods; D053719:Tandem Mass Spectrometry / Q000379:methods*
chemical_list: D002241:Carbohydrates; D011073:Polyamines; D000071228:Polyelectrolytes
keywords: Carbohydrates; Cyclodextrin; ESI-MS/MS; ETD; Gene delivery; Polycationic