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Título: | Revealing cooperative binding of polycationic cyclodextrins with DNA oligomers by capillary electrophoresis coupled to mass spectrometry |
Autor: | Przybylski, Cédric; Benito, Juan M. CSIC ORCID CVN ; Bonnet, Véronique; Ortiz-Mellet, Carmen; García Fernández, José Manuel CSIC ORCID | Palabras clave: | Gene delivery Hill number DNA Cooperative effects CE-MS Polycationic cyclodextrins |
Fecha de publicación: | 9-mar-2018 | Editor: | Elsevier | Citación: | Analytica Chimica Acta 1002: 70-81 (2018) | Resumen: | Gene delivery is critical for the development of nucleic acid-based therapies against a range of severe diseases. The conception of non-viral (semi)synthetic vectors with low cytotoxicity and virus-like efficiency is gathering a lot of efforts, but it represents a fantastic challenge still far from accomplishment. Carbohydrate-based scaffolds offer interesting features towards this end, such as easy availability, relatively cheap cost, tuning properties and a good biocompatibility. The lack of analytical methods providing quantitative and qualitative data on their binding properties with oligonucleotides (DNA/RNA), with a minimal time and sample consumption, represents a limitation for these channels. Here, we attempted to fill the gap by hyphenation of capillary electrophoresis with mass spectrometry (CE-MS). This coupling strategy allows discriminating free and complexed DNA oligomers with cationic cyclodextrins (CDs), determining the stoichiometry where the highest observed is always DNA: n/3(CD), and unambiguously assigning the partners through m/z detection. Very reliable data were obtained with migration time within 5.5 (standard deviation < 0.5%) and 25 min (standard deviation < 1.1%) for UV and MS detection, respectively. Furthermore, varying the nitrogen/phosphorus ratio (N/P), key parameters relating to the thermodynamics e.g. the micro and macroscopic dissociation constants K and K, respectively (both in low μM range) and the Gibbs free energy ΔG (−16.3 to −26.9 kJ mol), and also the cooperativity as Hill number (nH between 0.98 and 15.75) of the supramolecular process can be delineated, providing a unique tool for the high throughput screening and selection of efficient gene delivery carriers. | Versión del editor: | https://doi.org/10.1016/j.aca.2017.11.034 | URI: | http://hdl.handle.net/10261/190592 | DOI: | 10.1016/j.aca.2017.11.034 | ISSN: | 0003-2670 |
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