Diphtheria toxoid conformation in the context of its nanoencapsulation within liposomal particles sandwiched by chitosan

Diphtheria toxoid conformation in the context of its nanoencapsulation within liposomal particles sandwiched by chitosan

Author Rescia, Vanessa C. Autor UNIFESP Google Scholar
Ramos, Henrique R. Google Scholar
Takata, Celia S. Google Scholar
Araujo, Pedro S. de Google Scholar
Costa, Maria H. B. da Google Scholar
Institution Ctr Biotecnol
Div Desenvolvimento Tecnol & Prod
Universidade de São Paulo (USP)
Universidade Federal de São Paulo (UNIFESP)
Abstract Chitosan (alpha alpha-(1-4)-amino-2-deoxy-beta beta-D-glucan) is a deacetylated form of chitin, a polysaccharide from crustacean shells. Its unique characteristics, such as positive charge, biodegradability, biocompatibility, nontoxicity, and rigid structure, make this macromolecule ideal for an oral vaccine delivery system. We prepared reverse-phase evaporation vesicles (REVs) sandwiched by chitosan (Chi) and polyvinylic alcohol (PVA). However, in this method, there are still some problems to be circumvented related to protein stabilization. During the inverted micelle phase of protein nanoencapsulation, hydrophobic interfaces are expanded, leading to interfacial adsorption, followed by protein unfolding and aggregation. Here, spectroscopic and immunological techniques were used to ascertain the effects of the Hoffmeister series ions on diphtheria toxoid (Dtxd) stability during the inverted micelle phase. A correlation was established between the salts used in aqueous solutions and the changes in Dtxd solubility and conformation. Dtxd alpha alpha-helical content was quite stable, which led us to conclude that encapsulation occurred without protein aggregation or without exposition of hydrophobic residues. Dtxd aggregation was 98% avoided by the kosmotropic, PO<SU2--</SU(4). This ion was used to prepare a stable Dtxd and immunologically recognized REV-Chi-PVA formulation in the presence of 50 mM of PO<SU2--</SU(4). Under these conditions, the Dtxd retained its immunological identity. Therefore, we could obtain the maximum Dtxd solubility and stability after contact with CH(3)CO(2)C(2)H(5) to begin its nanoencapsulation within ideal conditions. This was a technological breakthrough, because a simple solution, such as salt, addition avoided heterologous protein use.</.
Keywords Liposome-vaccine delivery
Hoffmeister serie ions
protein stabilization
interaction protein/organic solvent
protein nanoencapsulation
Language English
Sponsor Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)
Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)
Fundacao Butantan
Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)
Grant number FAPESP: 00/10970-7
CNPq: 302047/2008-5
CNPq: 454177/2009-7
Date 2011-06-01
Published in Journal of Liposome Research. London: Informa Healthcare, v. 21, n. 2, p. 116-123, 2011.
ISSN 0898-2104 (Sherpa/Romeo, impact factor)
Publisher Informa Healthcare
Extent 116-123
Origin http://dx.doi.org/10.3109/08982104.2010.491072
Access rights Closed access
Type Article
Web of Science ID WOS:000290734600003
URI http://repositorio.unifesp.br/handle/11600/33790

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