Immunized mice naturally process in silico-derived peptides from the nucleocapsid of SARS-CoV-2

BMC Microbiology

Research

Marcela Salazar-García¹  Berenice Illades-Aguiar²  Mónica Espinoza-Rojo³  Karen Cortés-Sarabia³  Amalia Vences-Velázquez³  Mario Aldair Campos-Ruíz³  Victor M. Luna-Pineda⁴  Mariana Romo-Castillo⁵  Oscar Del Moral-Hernández⁶ 

[1] Laboratorio de Biología del Desarrollo y Teratogénesis Experimental, Hospital Infantil de México Federico Gómez, Ciudad de México, México;Laboratorio de Biomedicina Molecular, Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Guerrero, Chilpancingo de los Bravo, Guerrero, México;Laboratorio de Inmunobiología y Diagnóstico Molecular, Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Guerrero, Chilpancingo de los Bravo, Guerrero, México;Laboratorio de Investigación en COVID-19, Laboratorio de Investigación en Inmunología y Proteómica, Hospital Infantil de México Federico Gómez, Ciudad de México, México;Laboratorio de Investigación en COVID-19, Laboratorio de Investigación en Inmunología y Proteómica, Hospital Infantil de México Federico Gómez, Ciudad de México, México;Programa Investigadoras e Investigadores por México, Consejo Nacional de Humanidades, Ciencias y Tecnologías, Ciudad de México, México;Laboratorio de Virología, Facultad de Ciencias Químico Biológicas, Universidad Autónoma de Guerrero, Chilpancingo de los Bravo, Guerrero, México;

关键词: SARS-CoV-2;    Nucleocapsid protein;    Peptides;    In silico;    Immunoassays;   

DOI  :  10.1186/s12866-023-03076-5

 received in 2023-05-14, accepted in 2023-10-18,  发布年份 2023

来源: Springer

【 摘 要 】

BackgroundThe nucleocapsid (N) protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2) is an excellent immunogen that promotes the production of high-titer antibodies. N protein-derived peptides identified using a bioinformatics approach can potentially be used to develop a new generation of vaccines or diagnostic methods for detecting SARS-CoV-2 and its variants. However, further studies must demonstrate their capacity to be naturally processed by the immune system.ObjectiveWe aimed to examine the in vivo processing and recognition of in silico-identified peptides using the serum of immunized animals with the complete protein.MethodsRecombinant N (Nrec) protein was subcutaneously administered to six Balb/c mice. Enzyme-linked immunosorbent assay (ELISA), western blotting, dot blotting, and immunoprecipitation were performed to evaluate the recognition of the complete protein and in silico-derived peptides.ResultsThe serum of immunized mice recognized ~ 62.5 ng/µL of Nrec with high specificity to linear and conformational epitopes. Dot blot analysis showed that peptides Npep2 and Npep3 were the most reactive.ConclusionOur data confirm the high immunogenicity of the SARS-CoV-2 N protein and provide evidence on the antigenicity of two peptides located in the N-arm/RNA-binding domain (Npep2) and oligomerization domain/C-tail (Npep3), considered the biologically active site of the N protein.

【 授权许可】

CC BY   
© The Author(s) 2023

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