tetano
Editor, Senior Moderator
BMC Microbiol
. 2023 Oct 28;23(1):319.
doi: 10.1186/s12866-023-03076-5. Immunized mice naturally process in silico-derived peptides from the nucleocapsid of SARS-CoV-2
Mario Aldair Campos-Ruíz[SUP] 1 [/SUP], Berenice Illades-Aguiar[SUP] 2 [/SUP], Oscar Del Moral-Hernández[SUP] 3 [/SUP], Mariana Romo-Castillo[SUP] 4 5 [/SUP], Marcela Salazar-García[SUP] 6 [/SUP], Mónica Espinoza-Rojo[SUP] 1 [/SUP], Amalia Vences-Velázquez[SUP] 1 [/SUP], Karen Cortés-Sarabia[SUP] 7 [/SUP], Victor M Luna-Pineda[SUP] 8 [/SUP]
Affiliations
Background: The 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.
Objective: We aimed to examine the in vivo processing and recognition of in silico-identified peptides using the serum of immunized animals with the complete protein.
Methods: Recombinant 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.
Results: The 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.
Conclusion: Our 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.
Keywords: Immunoassays; In silico; Nucleocapsid protein; Peptides; SARS-CoV-2.
. 2023 Oct 28;23(1):319.
doi: 10.1186/s12866-023-03076-5. Immunized mice naturally process in silico-derived peptides from the nucleocapsid of SARS-CoV-2
Mario Aldair Campos-Ruíz[SUP] 1 [/SUP], Berenice Illades-Aguiar[SUP] 2 [/SUP], Oscar Del Moral-Hernández[SUP] 3 [/SUP], Mariana Romo-Castillo[SUP] 4 5 [/SUP], Marcela Salazar-García[SUP] 6 [/SUP], Mónica Espinoza-Rojo[SUP] 1 [/SUP], Amalia Vences-Velázquez[SUP] 1 [/SUP], Karen Cortés-Sarabia[SUP] 7 [/SUP], Victor M Luna-Pineda[SUP] 8 [/SUP]
Affiliations
- PMID: 37898784
- DOI: 10.1186/s12866-023-03076-5
Background: The 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.
Objective: We aimed to examine the in vivo processing and recognition of in silico-identified peptides using the serum of immunized animals with the complete protein.
Methods: Recombinant 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.
Results: The 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.
Conclusion: Our 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.
Keywords: Immunoassays; In silico; Nucleocapsid protein; Peptides; SARS-CoV-2.