Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response
Introduction: The SARS-CoV-2 pandemic necessitates effective therapeutic solutions. The receptor-binding motif (RBM) is a subdomain of the spike protein's receptor-binding domain (RBD) and is critical for facilitating the binding of SARS-CoV-2 to the human ACE2 receptor. This study investigates...
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Tabriz University of Medical Sciences
2024-11-01
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| Online Access: | https://bi.tbzmed.ac.ir/PDF/bi-15-30520.pdf |
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| author | Hossein Samiei-Abianeh Shahram Nazarian Emad Kordbacheh Alireza Felegary |
| author_facet | Hossein Samiei-Abianeh Shahram Nazarian Emad Kordbacheh Alireza Felegary |
| author_sort | Hossein Samiei-Abianeh |
| collection | DOAJ |
| description | Introduction: The SARS-CoV-2 pandemic necessitates effective therapeutic solutions. The receptor-binding motif (RBM) is a subdomain of the spike protein's receptor-binding domain (RBD) and is critical for facilitating the binding of SARS-CoV-2 to the human ACE2 receptor. This study investigates the use of the receptor-binding motif (RBM) domain as an immunogen to produce potent neutralizing antibodies against SARS-CoV-2. Methods: The RBM gene was codon-optimized and cloned into the pET17b vector for expression in E. coli BL21 (DE3) cells, induced with 1 mM IPTG. The recombinant RBM protein was purified using Ni-NTA affinity chromatography. After validating the recombinant RBM by Western blotting with anti-His tag antibodies, BALB/c mice were immunized with 20 µg of the purified RBM protein. Anti-RBM IgG was subsequently purified using protein G resin, and its neutralizing capacity was assessed using the Pishtaz Teb Zaman Neutralization Assay Kit. Results: The recombinant RBM protein, with a molecular weight of 10 kDa, was expressed as inclusion bodies. the typical yield of purification was 27 mg/L of bacterial culture. The neutralization test demonstrated a concentration of 36 µg/mL of neutralizing antibodies in the immunized serum, preventing the spike protein from binding to ACE2. Conclusion: Our study demonstrated that anti-RBM antibodies exhibited neutralization effects on SARS-CoV-2. These findings provide evidence for the development of a vaccine candidate through the induction of antibodies against the RBM, necessitating further studies with adjuvants suitable for human use to evaluate its potential for human vaccination. |
| format | Article |
| id | doaj-art-af3b2b2138f24aed87157f829bb2d5cd |
| institution | DOAJ |
| issn | 2228-5652 2228-5660 |
| language | English |
| publishDate | 2024-11-01 |
| publisher | Tabriz University of Medical Sciences |
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| series | BioImpacts |
| spelling | doaj-art-af3b2b2138f24aed87157f829bb2d5cd2025-08-20T02:57:54ZengTabriz University of Medical SciencesBioImpacts2228-56522228-56602024-11-01151305203052010.34172/bi.30520bi-30520Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody responseHossein Samiei-Abianeh0Shahram Nazarian1Emad Kordbacheh2Alireza Felegary3Department of Medical Biotechnology and Nanotechnology, Faculty of Medicine, Mashhad University of Medical Sciences, Mashhad, IranDepartment of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, IranDepartment of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, IranDepartment of Biology, Faculty of Basic Sciences, Imam Hossein University, Tehran, IranIntroduction: The SARS-CoV-2 pandemic necessitates effective therapeutic solutions. The receptor-binding motif (RBM) is a subdomain of the spike protein's receptor-binding domain (RBD) and is critical for facilitating the binding of SARS-CoV-2 to the human ACE2 receptor. This study investigates the use of the receptor-binding motif (RBM) domain as an immunogen to produce potent neutralizing antibodies against SARS-CoV-2. Methods: The RBM gene was codon-optimized and cloned into the pET17b vector for expression in E. coli BL21 (DE3) cells, induced with 1 mM IPTG. The recombinant RBM protein was purified using Ni-NTA affinity chromatography. After validating the recombinant RBM by Western blotting with anti-His tag antibodies, BALB/c mice were immunized with 20 µg of the purified RBM protein. Anti-RBM IgG was subsequently purified using protein G resin, and its neutralizing capacity was assessed using the Pishtaz Teb Zaman Neutralization Assay Kit. Results: The recombinant RBM protein, with a molecular weight of 10 kDa, was expressed as inclusion bodies. the typical yield of purification was 27 mg/L of bacterial culture. The neutralization test demonstrated a concentration of 36 µg/mL of neutralizing antibodies in the immunized serum, preventing the spike protein from binding to ACE2. Conclusion: Our study demonstrated that anti-RBM antibodies exhibited neutralization effects on SARS-CoV-2. These findings provide evidence for the development of a vaccine candidate through the induction of antibodies against the RBM, necessitating further studies with adjuvants suitable for human use to evaluate its potential for human vaccination.https://bi.tbzmed.ac.ir/PDF/bi-15-30520.pdfcovid-19spike glycoproteinreceptor-binding domainreceptor-binding motifrecombinant proteinsubunit vaccine |
| spellingShingle | Hossein Samiei-Abianeh Shahram Nazarian Emad Kordbacheh Alireza Felegary Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response BioImpacts covid-19 spike glycoprotein receptor-binding domain receptor-binding motif recombinant protein subunit vaccine |
| title | Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response |
| title_full | Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response |
| title_fullStr | Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response |
| title_full_unstemmed | Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response |
| title_short | Recombinant receptor-binding motif of spike COVID-19 vaccine candidate induces SARS-CoV-2 neutralizing antibody response |
| title_sort | recombinant receptor binding motif of spike covid 19 vaccine candidate induces sars cov 2 neutralizing antibody response |
| topic | covid-19 spike glycoprotein receptor-binding domain receptor-binding motif recombinant protein subunit vaccine |
| url | https://bi.tbzmed.ac.ir/PDF/bi-15-30520.pdf |
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