دورية أكاديمية
Obtaining a high titer of polyclonal antibodies from rats to the SARS-CoV-2 nucleocapsid protein and its N- and C-terminal domains for diagnostic test development.
| العنوان: | Obtaining a high titer of polyclonal antibodies from rats to the SARS-CoV-2 nucleocapsid protein and its N- and C-terminal domains for diagnostic test development. |
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| المؤلفون: | de Almeida MT; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Barbosa AP; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Bomfim CG; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Visnardi AB; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Vinces TC; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Ceroni A; Department of Physiology and Biophysics, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil., Durigon EL; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil; Scientific Platform Pasteur USP, São Paulo, Brazil., Guzzo CR; Department of Microbiology, Institute of Biomedical Sciences, University of São Paulo, São Paulo, Brazil. Electronic address: crisguzzo@usp.br. |
| المصدر: | Journal of immunological methods [J Immunol Methods] 2023 Nov; Vol. 522, pp. 113558. Date of Electronic Publication: 2023 Sep 11. |
| نوع المنشور: | Journal Article; Research Support, Non-U.S. Gov't |
| اللغة: | English |
| بيانات الدورية: | Publisher: Elsevier Country of Publication: Netherlands NLM ID: 1305440 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1872-7905 (Electronic) Linking ISSN: 00221759 NLM ISO Abbreviation: J Immunol Methods Subsets: MEDLINE |
| أسماء مطبوعة: | Publication: Amsterdam : Elsevier Original Publication: Amsterdam, North-Holand, |
| مواضيع طبية MeSH: | SARS-CoV-2*/genetics , COVID-19*/diagnosis, Humans ; Female ; Animals ; Rats ; Antibodies, Viral ; COVID-19 Vaccines ; Rats, Wistar ; Antibodies, Neutralizing ; Nucleocapsid Proteins/genetics ; Diagnostic Tests, Routine ; COVID-19 Testing |
| مستخلص: | Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) is an enveloped, plus-stranded RNA virus responsible for the Coronavirus Disease 2019 (COVID-19). Patients infected with COVID-19 may be asymptomatic or have symptoms ranging from mild manifestations to severe cases of the disease that could lead to death. The SARS-CoV-2 genome encodes 4 structural proteins, including the Spike protein (S), the Nucleocapsid protein (N), Membrane protein (M) and, the Envelope protein (E). The N protein forms a major component of the ribonucleoprotein complex within the virus particle and play a vital role in its transcription and replication. Nevertheless, the S protein was the most important protein in the development of vaccines against COVID-19. However, the decrease in number of registered immunizations against the disease and the rapid drop in neutralizing antibody titers together with looser preventive measures for virus transmission, favored the rapid appearance of new variants of concerns (VOCs) that primarily show mutations in the S protein. This fact makes the N protein a good candidate for the development of diagnostic tests, due to its stability, amino acid conservation, high immunogenicity, and the smaller likelihood of mutation. With the aim of developing a new diagnostic kit based on the N protein, we evaluated the humoral response in female Wistar rats against this target. Three constructions of the N protein were used to inoculate the animals: the full-length protein (Cfull), the N- (NTD), and the C-terminal (CTD) portion of the protein. The immunizations induced the animal's immune response, with specific polyclonal IgG antibodies against the Cfull protein and its fragments. There were not non-specific bind to the protein used as negative control. Anti-Cfull antibodies demonstrated high efficiency in binding to the NTD protein and the antibodies present in the anti-CTD and anti-NTD sera have recognized the Cfull protein, but they were not able to recognize the NTD and CTD proteins, respectively. Our results indicate an efficient protocol for obtaining high antibody titers against the N recombinant protein of SARS-CoV-2 and its fragments highlighting the Cfull protein, which can be used in the development of new diagnostic kits. Competing Interests: Declaration of Competing Interest The authors declare no conflict of interest. (Copyright © 2023. Published by Elsevier B.V.) |
| فهرسة مساهمة: | Keywords: Antibodies; COVID-19; Conserved protein; Diagnosis; Nucleocapsid |
| المشرفين على المادة: | 0 (Antibodies, Viral) 0 (COVID-19 Vaccines) 0 (Antibodies, Neutralizing) 0 (Nucleocapsid Proteins) |
| تواريخ الأحداث: | Date Created: 20230913 Date Completed: 20231030 Latest Revision: 20231031 |
| رمز التحديث: | 20240628 |
| DOI: | 10.1016/j.jim.2023.113558 |
| PMID: | 37704125 |
| قاعدة البيانات: | MEDLINE |
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