Performance of protein N linear epitopes in serodiagnosis of COVID-19 infection

Author:

Farajnia Sahar12ORCID,Khajenasiri Nazli1,Farajnia Safar23ORCID,Seyrafi Farzin3,Bakhtiyari Nasim34

Affiliation:

1. Department of Biology, University of Rabe Rashidi, Tabriz, Iran

2. Drug Applied Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

3. Biotechnology Research Center, Tabriz University of Medical Sciences, Tabriz, Iran

4. Student Research Committee, Tabriz University of Medical Sciences, Tabriz, Iran

Abstract

Introduction: Despite the efforts to contain the spread of COVID-19, the virus remains in circulation, posing a considerable risk to populations across the globe. Hence, rapid and early detection of this infection is essential for effective disease control. The nucleocapsid (N) protein of the virus serves as a primary target for antibody response during CoV2 infections, making it a potential candidate for COVID-19 detection. This study aims to prepare and evaluate the linear epitopes of the N protein for serodiagnosis of COVID-19 infection. Methods: The linear epitope of the N protein gene was identified using ABCpred, BCpred, and IEDB. These epitopes were subsequently amplified by RT-PCR, cloned, and expressed in soluble form in the E. coli BL21 strain. The recombinant protein was purified using the Ni-NTA column. The reactivity of purified N protein with sera from SARS-CoV-2 patients was analyzed using an ELISA assay. Results: Sequencing analysis demonstrated the successful cloning of the linear epitopes of the N protein into the PET-28a vector, along with an n-terminal His-tag fusion. The recombinant protein was produced in E. coli BL21 and purified with a Ni-NTA column. The analysis demonstrated that the N protein linear epitopes were expressed in a soluble form and appeared as a 50 kDa band in the SDS-PAGE. Examination for the reactivity of the purified N protein with the COVID-19 patient’s sera by ELISA revealed that the N protein recognizes the infection with high sensitivity and specificity. Conclusion: The results of this study indicated that linear epitopes of the SARS-CoV-2 N protein are highly immunogenic and could be exploited for serodiagnosis of infection in patients suspected of COVID-19 infection.

Publisher

Maad Rayan Publishing Company

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