Search

Your search keyword '"Rodríguez-Mallón A"' showing total 2 results
Start Over Author "Rodríguez-Mallón A" Topic biochemistry
2 results on '"Rodríguez-Mallón A"'

1. Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein

Author

Raúl Mejías, Rafmary Rodríguez Fernández, Gleysin Cabrera, Pedro E. Encinosa Guzmán, Jose L Alvarez, Hilda Garay, Gervásio Henrique Bechara, Alina Rodríguez Mallón, Ania Cabrales, Madelón Portela, Yamil Bello Soto, Mario Pablo Estrada, Alier Fuentes Castillo, Alexei F. Licea-Navarro, Samanta Jiménez, Osmany Guirola, Luis Méndez, Gustavo Seron Sanches, Luis Javier González, Rosario Durán, Fabiola Almeida, Satomy Pousa, Center for Genetic Engineering and Biotechnology (CIGB), CIGB, Pontifícia Universidade Católica do Paraná (PUCPR), Universidade Estadual Paulista (Unesp), Instituto de Ciencia Animal (ICA), National Laboratory for Parasitology, CICESE, Institut Pasteur de Montevideo, and Instituto de Investigaciones Biológicas Clemente Estable (IIBCE)

Subjects
0301 basic medicine, Microbiology (medical), Immunogen, Bm86, chemical conjugation, Rhipicephalus sanguineus, 030231 tropical medicine, lcsh:Medicine, Peptide, anti-tick vaccine, Megathura crenulata, Article, Anti-tick vaccine, 03 medical and health sciences, 0302 clinical medicine, Antigen, parasitic diseases, Immunology and Allergy, Molecular Biology, cross-linked peptides, chemistry.chemical_classification, General Immunology and Microbiology, biology, Immunogenicity, lcsh:R, Cross-linked peptides, Chemical conjugation, P0, biology.organism_classification, peptide, 030104 developmental biology, Infectious Diseases, chemistry, Biochemistry, biology.protein, Rhipicephalus microplus, Keyhole limpet hemocyanin
Abstract

A synthetic 20 amino acid peptide of the ribosomal protein P0 from ticks, when conjugated to keyhole limpet hemocyanin from Megathura crenulata and used as an immunogen against Rhipicephalus microplus and Rhipicephalus sanguineus s.l. species, has shown efficacies of around 90%. There is also experimental evidence of a high efficacy of this conjugate against Amblyomma mixtum and Ixodes ricinus species, which suggest that this antigen could be a good broad-spectrum anti-tick vaccine candidate. In this study, the P0 peptide (pP0) was chemically conjugated to Bm86 as a carrier protein. SDS-PAGE analysis of this conjugate demonstrated that it is highly heterogeneous in size, carrying from 1 to 18 molecules of pP0 per molecule of Bm86. Forty-nine out of the 54 lysine residues and the N-terminal end of Bm86 were found partially linked to pP0 by using LC-MS/MS analysis and the combination of four different softwares. Several post-translational modifications of Bm86 protein were also identified by mass spectrometry. High immunogenicity and efficacy were achieved when dogs and cattle were vaccinated with the pP0&ndash, Bm86 conjugate and challenged with R. sanguineus s.l. and R. microplus, respectively. These results encourage the development of this antigen with promising possibilities as an anti-tick vaccine.

Published
2020
Full Text
View/download PDF

2. Conformational and Thermal Stability Improvements for the Large-Scale Production of Yeast-Derived Rabbit Hemorrhagic Disease Virus-Like Particles as Multipurpose Vaccine.

Author

Fernández, Erlinda, Toledo, Jorge R., Méndez, Lídice, González, Nemecio, Parra, Francisco, Martín-Alonso, José M., Limonta, Miladys, Sánchez, Kosara, Cabrales, Ania, Estrada, Mario P., Rodríguez-Mallón, Alina, and Farnós, Omar

Subjects
RABBIT calicivirus disease, VETERINARY vaccines, THERMAL stability, CONFORMATIONAL analysis, BIOTECHNOLOGY research, VIRAL vaccines
Abstract

Recombinant virus-like particles (VLP) antigenically similar to rabbit hemorrhagic disease virus (RHDV) were recently expressed at high levels inside Pichia pastoris cells. Based on the potential of RHDV VLP as platform for diverse vaccination purposes we undertook the design, development and scale-up of a production process. Conformational and stability issues were addressed to improve process control and optimization. Analyses on the structure, morphology and antigenicity of these multimers were carried out at different pH values during cell disruption and purification by size-exclusion chromatography. Process steps and environmental stresses in which aggregation or conformational instability can be detected were included. These analyses revealed higher stability and recoveries of properly assembled high-purity capsids at acidic and neutral pH in phosphate buffer. The use of stabilizers during long-term storage in solution showed that sucrose, sorbitol, trehalose and glycerol acted as useful aggregation-reducing agents. The VLP emulsified in an oil-based adjuvant were subjected to accelerated thermal stress treatments. None to slight variations were detected in the stability of formulations and in the structure of recovered capsids. A comprehensive analysis on scale-up strategies was accomplished and a nine steps large-scale production process was established. VLP produced after chromatographic separation protected rabbits against a lethal challenge. The minimum protective dose was identified. Stabilized particles were ultimately assayed as carriers of a foreign viral epitope from another pathogen affecting a larger animal species. For that purpose, a linear protective B-cell epitope from Classical Swine Fever Virus (CSFV) E2 envelope protein was chemically coupled to RHDV VLP. Conjugates were able to present the E2 peptide fragment for immune recognition and significantly enhanced the peptide-specific antibody response in vaccinated pigs. Overall these results allowed establishing improved conditions regarding conformational stability and recovery of these multimers for their production at large-scale and potential use on different animal species or humans. [ABSTRACT FROM AUTHOR]

Published
2013
Full Text
View/download PDF

Catalog

Books, media, physical & digital resources
See catalog results