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Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry

Highly branched polyamidoamine (PAMAM) dendrimers presenting biological activities have been envisaged as non-viral gene delivery vectors. They are known to associate with nucleic acid (DNA) in non-covalent complexes via electrostatic interactions. Although their transfection efficiency has been pro...

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Autores principales: Leriche, Emma-Dune, Hubert-Roux, Marie, Afonso, Carlos, Lange, Catherine M., Grossel, Martin C., Maire, Florian, Loutelier-Bourhis, Corinne
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6271531/
https://www.ncbi.nlm.nih.gov/pubmed/25514219
http://dx.doi.org/10.3390/molecules191220731
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author Leriche, Emma-Dune
Hubert-Roux, Marie
Afonso, Carlos
Lange, Catherine M.
Grossel, Martin C.
Maire, Florian
Loutelier-Bourhis, Corinne
author_facet Leriche, Emma-Dune
Hubert-Roux, Marie
Afonso, Carlos
Lange, Catherine M.
Grossel, Martin C.
Maire, Florian
Loutelier-Bourhis, Corinne
author_sort Leriche, Emma-Dune
collection PubMed
description Highly branched polyamidoamine (PAMAM) dendrimers presenting biological activities have been envisaged as non-viral gene delivery vectors. They are known to associate with nucleic acid (DNA) in non-covalent complexes via electrostatic interactions. Although their transfection efficiency has been proved, PAMAMs present a significant cytotoxicity due to their cationic surface. To overcome such a drawback, different chemical modifications of the PAMAM surface have been reported such as the attachment of hydrophobic residues. In the present work, we studied the complexation of DNA duplexes with different low-generation PAMAM; ammonia-cored G0(N) and G1(N) PAMAM, native or chemically modified with aromatic residues, i.e., phenyl-modified-PAMAM G0(N) and phenylalanine-modified-PAMAM G1(N). To investigate the interactions involved in the PAMAM/DNA complexes, also called dendriplexes, we used electrospray ionization (ESI) coupled to ion mobility spectrometry-mass-spectrometry (IM-MS). ESI is known to allow the study of non-covalent complexes in native conditions while IM-MS is a bidimensional separation technique particularly useful for the characterization of complex mixtures. IM-MS allows the separation of the expected complexes, possible additional non-specific complexes and the free ligands. Tandem mass spectrometry (MS/MS) was also used for the structural characterization. This work highlights the contribution of IM-MS and MS/MS for the study of small dendriplexes. The stoichiometries of the complexes and the equilibrium dissociation constants were determined. The [DNA/native PAMAM] and [DNA/modified-PAMAM] dendriplexes were compared.
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spelling pubmed-62715312018-12-28 Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry Leriche, Emma-Dune Hubert-Roux, Marie Afonso, Carlos Lange, Catherine M. Grossel, Martin C. Maire, Florian Loutelier-Bourhis, Corinne Molecules Article Highly branched polyamidoamine (PAMAM) dendrimers presenting biological activities have been envisaged as non-viral gene delivery vectors. They are known to associate with nucleic acid (DNA) in non-covalent complexes via electrostatic interactions. Although their transfection efficiency has been proved, PAMAMs present a significant cytotoxicity due to their cationic surface. To overcome such a drawback, different chemical modifications of the PAMAM surface have been reported such as the attachment of hydrophobic residues. In the present work, we studied the complexation of DNA duplexes with different low-generation PAMAM; ammonia-cored G0(N) and G1(N) PAMAM, native or chemically modified with aromatic residues, i.e., phenyl-modified-PAMAM G0(N) and phenylalanine-modified-PAMAM G1(N). To investigate the interactions involved in the PAMAM/DNA complexes, also called dendriplexes, we used electrospray ionization (ESI) coupled to ion mobility spectrometry-mass-spectrometry (IM-MS). ESI is known to allow the study of non-covalent complexes in native conditions while IM-MS is a bidimensional separation technique particularly useful for the characterization of complex mixtures. IM-MS allows the separation of the expected complexes, possible additional non-specific complexes and the free ligands. Tandem mass spectrometry (MS/MS) was also used for the structural characterization. This work highlights the contribution of IM-MS and MS/MS for the study of small dendriplexes. The stoichiometries of the complexes and the equilibrium dissociation constants were determined. The [DNA/native PAMAM] and [DNA/modified-PAMAM] dendriplexes were compared. MDPI 2014-12-12 /pmc/articles/PMC6271531/ /pubmed/25514219 http://dx.doi.org/10.3390/molecules191220731 Text en © 2014 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Leriche, Emma-Dune
Hubert-Roux, Marie
Afonso, Carlos
Lange, Catherine M.
Grossel, Martin C.
Maire, Florian
Loutelier-Bourhis, Corinne
Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry
title Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry
title_full Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry
title_fullStr Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry
title_full_unstemmed Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry
title_short Investigation of Dendriplexes by Ion Mobility-Mass Spectrometry
title_sort investigation of dendriplexes by ion mobility-mass spectrometry
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6271531/
https://www.ncbi.nlm.nih.gov/pubmed/25514219
http://dx.doi.org/10.3390/molecules191220731
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