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

Autor: Alina Rodríguez Mallón, Luis Javier González, Pedro Enrique Encinosa Guzmán, Gervasio Henrique Bechara, Gustavo Seron Sanches, Satomy Pousa, Gleysin Cabrera, Ania Cabrales, Hilda Garay, Raúl Mejías, José Raúl López Álvarez, Yamil Bello Soto, Fabiola Almeida, Osmany Guirola, Rafmary Rodríguez Fernández, Alier Fuentes Castillo, Luis Méndez, Samanta Jiménez, Alexei Licea-Navarro, Madelón Portela, Rosario Durán, Mario Pablo Estrada
Jazyk: angličtina
Rok vydání: 2020
Předmět:
Zdroj: Pathogens, Vol 9, Iss 6, p 513 (2020)
Druh dokumentu: article
ISSN: 2076-0817
DOI: 10.3390/pathogens9060513
Popis: 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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