Recombinant expression of SARS-CoV-2 Receptor Binding Domain (RBD) in Escherichia coli and its immunogenicity in mice | ||
| Iranian Journal of Basic Medical Sciences | ||
| مقاله 10، دوره 25، شماره 9، آذر 2022، صفحه 1110-1116 اصل مقاله (458.56 K) | ||
| نوع مقاله: Original Article | ||
| شناسه دیجیتال (DOI): 10.22038/ijbms.2022.65045.14333 | ||
| نویسندگان | ||
| zahra Rahbar1؛ Shahram Nazarian2؛ Ruhollah Dorostkar3؛ Fattah Sotoodehnejadnematalahi1؛ Jafar Amani* 4 | ||
| 1Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran | ||
| 2Department of Biology, Imam Hossein University, Tehran, Iran | ||
| 3Applied Virology Research Center, Baqiyatallah University of Medical Sciences, Iran | ||
| 4Applied Microbiology Research Center, System Biology and Poisonings Institute, Baqiyatallah University of Medical Sciences, Tehran, Iran | ||
| چکیده | ||
| Objective(s): The severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), giving rise to the coronavirus disease 2019 (COVID-19), has become a danger to wellbeing worldwide. Thus, finding efficient and safe vaccines for COVID-19 is of great importance. As a basic step amid contamination, SARS-CoV-2 employs the receptor-binding domain (RBD) of the spike protein to lock in with the receptor angiotensin-converting enzyme 2 (ACE2) on host cells. SARS-CoV-2 receptor-binding domain (RBD) is the main human antibody target for developing vaccines and virus inhibitors, as well as neutralizing antibodies. A bacterial procedure was developed for the expression and purification of the SARS-CoV-2 spike protein receptor-binding domain. Materials and Methods: In this research study, RBD was expressed by Escherichia coli and purified with Ni-NTA chromatography. Then it was affirmed by the western blot test. The immunogenicity and protective efficacy of RBD recombinant protein were assessed on BALB/c mice. Additionally, RBD recombinant protein was tested by ELISA utilizing sera of COVID-19 healing patients contaminated with SARS-CoV-2 wild type and Delta variation. Results: Indirect ELISA was able to detect the protein RBD in serum of the immunized mouse expressed in E. coli. The inactive SARS-CoV2 was detected by antibodies within the serum of immunized mice. Serum antibodies from individuals recovered from Covid19 reacted to the expressed protein. Conclusion: Our findings showed that RBD is of great importance in vaccine design and it can be used to develop recombinant vaccines through induction of antibodies against RBD. | ||
| کلیدواژهها | ||
| Neutralizing antibodies؛ RBD؛ Recombinant proteins؛ SARS-CoV-2؛ Vaccines | ||
| مراجع | ||
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