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:: Volume 24, Issue 6 (November and December 2022 2022) ::
Koomesh 2022, 24(6): 727-735 Back to browse issues page
Evaluation of expression and titration of recombinant nucleocapsid protein as an immunogenic candidate against SARS-CoV2
Hossein Samiei Abianeh , Shahram Nazarian , Jafar Amani , Emad Kordbache , Amir Rezaie
Abstract:   (552 Views)
Introduction: Covid-19 epidemic results from an infection caused by SARS-CoV2. Evolution-based analyses on the nucleotide sequences show that SARS-CoV2 is a member of the genus Beta-coronaviruses and its genome consists of a single-stranded RNA, encoding 16 proteins. Among the structural proteins, the nucleocapsid is the most abundant protein in virus structure, highly immunogenic, with sequence conservatory. Due to a large number of mutations in the spike protein, the aim of this study was to investigate bioinformatics, expression of nucleocapsid protein and evaluate its immunogenicity as an immunogenic candidate
Materials and Methods: B and T cell epitopes of nucleocapsid protein were examined in the IEDB database. The PET28a-N plasmid was transferred to E. coli BL21(DE3) expression host, and IPTG induced recombinant protein expression. The protein was purified using Ni-NTA column affinity chromatography, and the Western blotting method was utilized to confirm it. Finally, mice were immunized with three routes of purified protein. Statistical analysis of the control group injection and test results was carried out by t-test from SPSS software.
Results: The optimized gene had a Codon adaptation index (CAI) of 0/97 Percentage of codons having high- frequency distribution was improved to 85%. Expression of recombinant protein in E.coli led to the production of BoNT/B-HCC with a molecular weight of 45 kDa. The total yield of purified protein was 43 mg/L. Immunization of mice induced serum antibody response. Statistical analysis showed that the antibody titer ratio was significantly different compared to the control sample and the antibody titer was acceptable up to a dilution of 1.256000
Conclusion: According to the present study results, the protein can be used as an immunogenic candidate for developing vaccines against SARS-CoV2 in future research.
Keywords: Covid-19, SARS-CoV2, Nucleocapsid, Recombinant Immunogenic, Epitope
Full-Text [PDF 1578 kb]   (346 Downloads)    
Type of Study: Research | Subject: General
Received: 2022/02/9 | Accepted: 2022/06/22 | Published: 2022/11/28
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Samiei Abianeh H, Nazarian S, Amani J, Kordbache E, Rezaie A. Evaluation of expression and titration of recombinant nucleocapsid protein as an immunogenic candidate against SARS-CoV2. Koomesh 2022; 24 (6) :727-735
URL: http://koomeshjournal.semums.ac.ir/article-1-7510-en.html


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Volume 24, Issue 6 (November and December 2022 2022) Back to browse issues page
کومش Koomesh
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