Expression and Purification of SARS CoV-2 RBD Protein in the Prokaryotic Expression System to Evaluate the IgG Antibody in the Serums of COVID-19 Recovered Cases

Document Type : Original Article

Authors

1 Department of Virology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Department of Influenza and Respiratory Diseases, Pasteur Institute, Tehran, Iran

3 Hepatitis and AIDS Department, Pasteur Institute, Tehran, Iran

10.61186/jsmj.2022.341969.2843

Abstract

Background and Objectives One of the most important proteins of the SARS-COV-
2 virus is S protein, which consists of S1 and S2 subunits. The most important region in
subunit S1 is the receptor binding region (RBD), which plays a key role in binding to
ACE receptors. The RBD is a conserved region in S protein that is the target of the
immune system and the production of antibodies against the virus. Due to the
importance of this region in the production of neutralizing antibodies, it can be a good
candidate for vaccine development and production of diagnostic kits. Therefore, the
present study aimed to assess the response rate of antibodies in the serum of
recovered COVID-19 cases using recombinant RBD protein produced in the prokaryotic
expression system as an inexpensive expression system.
Subjects and Methods In this study, the recombinant PET22b-RBD construct was
transformed to the Escherichia coli (BL21) host, and the bacterial cells were cultured
in a culture medium; thereafter, protein expression was induced using isopropyl beta-
D-thiogalactoside (IPTG). The expression of recombinant RBD protein was confirmed
by acrylamide gel (SDS-PAGE) and Western blotting. Finally, the desired protein was
extracted using a Ni-NTA column and applied in an indirect ELISA.
Results In comparison with the serum of healthy individuals (cut off: 0.412), no
significant increase was observed in the response of 30 serum samples to the
recombinant RBD protein.
Conclusion The recombinant RBD protein produced in the prokaryotic host did not
respond significantly to the antibodies in the serum of recovered COVID-19 patients
and cannot be used for diagnostic purposes.

Keywords

Main Subjects

References

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Jundishapur Scientific Medical Journal
Volume 22, Issue 1 - Serial Number 142
March and April 2023
Pages 1-10

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History

  • Receive Date: 12 May 2022
  • Revise Date: 12 July 2022
  • Accept Date: 20 July 2022

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