Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein
by
Alina Rodríguez Mallón 1,*
, Luis Javier González 2, 
Pedro Enrique Encinosa Guzmán 1, Gervasio Henrique Bechara 3,4,†, Gustavo Seron Sanches 3,4,†, Satomy Pousa 2, Gleysin Cabrera 2, Ania Cabrales 5, Hilda Garay 5, Raúl Mejías 6, José Raúl López Álvarez 6, Yamil Bello Soto 1, Fabiola Almeida 2, Osmany Guirola 7, Rafmary Rodríguez Fernández 8, Alier Fuentes Castillo 8, Luis Méndez 8, Samanta Jiménez 9, Alexei Licea-Navarro 9, Madelón Portela 10,11, Rosario Durán 10,11 and Mario Pablo Estrada 1
Alina Rodríguez Mallón 1,*, Luis Javier González 2, Pedro Enrique Encinosa Guzmán 1, Gervasio Henrique Bechara 3,4,†, Gustavo Seron Sanches 3,4,†, Satomy Pousa 2, Gleysin Cabrera 2, Ania Cabrales 5, Hilda Garay 5, Raúl Mejías 6, José Raúl López Álvarez 6, Yamil Bello Soto 1, Fabiola Almeida 2, Osmany Guirola 7, Rafmary Rodríguez Fernández 8, Alier Fuentes Castillo 8, Luis Méndez 8, Samanta Jiménez 9, Alexei Licea-Navarro 9, Madelón Portela 10,11, Rosario Durán 10,11 and Mario Pablo Estrada 1
1
Animal Biotechnology Department, Center for Genetic Engineering and Biotechnology (CIGB), Havana 10600, Cuba
2
Mass Spectrometry Laboratory and GlycoLab, Department of Proteomics, CIGB, Havana 10600, Cuba
3
Programa de Pós-graduação em Ciência Animal, Pontifícia Universidade Católica do Paraná (PUCPR), Paraná 80215-901, Brazil
4
Departamento de Patologia Veterinária, Faculdade de Ciências Agrárias e Veterinárias, Universidade Estadual Paulista (FCAV-UNESP), São Paulo 14884-900, Brazil
5
Synthetic Peptides Group, CIGB, Havana 10600, Cuba
6
Instituto de Ciencia Animal (ICA), San José de las Lajas 32700, Cuba
7
Bioinformatic Department, CIGB, Havana 10600, Cuba
8
National Laboratory for Parasitology, San Antonio de los Banos 32500, Cuba
9
Departamento de Innovación Biomédica, CICESE, Ensenada 22860, Mexico
10
Unidad de Bioquímica y Proteómica Analítica, Institut Pasteur de Montevideo, Montevideo 11400, Uruguay
11
Instituto de Investigaciones Biológicas Clemente Estable (IIBCE), Montevideo 11600, Uruguay
*
Author to whom correspondence should be addressed.
†
Present address of Prof. Gervasio H. Bechara and Gustavo S. Sanches.
Pathogens 2020, 9(6), 513; https://doi.org/10.3390/pathogens9060513
Received: 3 June 2020 / Revised: 20 June 2020 / Accepted: 21 June 2020 / Published: 25 June 2020
(This article belongs to the Special Issue New Frontiers in Tick Research)
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–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.
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Keywords:
anti-tick vaccine; P0; Bm86; peptide; chemical conjugation; cross-linked peptides
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MDPI and ACS Style
Rodríguez Mallón, A.; Javier González, L.; Encinosa Guzmán, P.E.; Bechara, G.H.; Sanches, G.S.; Pousa, S.; Cabrera, G.; Cabrales, A.; Garay, H.; Mejías, R.; López Álvarez, J.R.; Bello Soto, Y.; Almeida, F.; Guirola, O.; Rodríguez Fernández, R.; Fuentes Castillo, A.; Méndez, L.; Jiménez, S.; Licea-Navarro, A.; Portela, M.; Durán, R.; Estrada, M.P. Functional and Mass Spectrometric Evaluation of an Anti-Tick Antigen Based on the P0 Peptide Conjugated to Bm86 Protein. Pathogens 2020, 9, 513.
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