Evaluation of expression and titration of recombinant nucleocapsid protein as an immunogenic candidate against SARS-CoV2
Koomesh
; 24(6), 2022.
Article
Dans Persan
| CAB Abstracts | ID: covidwho-20231716
ABSTRACT
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 andMethods:
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.
Prion; Viral; Bacterial and Fungal Pathogens of Humans [VV210]; Genetics and Molecular Biology of Microorganisms [ZZ395]; Animal Models of Human Diseases [VV400]; Host Resistance and Immunity [HH600]; Animal and in vitro Models for Pharmaceuticals [VV450]; genes; gene expression; antigens; proteins; human diseases; viral diseases; coronavirus disease 2019; pandemics; nucleocapsid proteins; titration; epidemics; viral proteins; epitopes; T lymphocytes; B lymphocytes; antibodies; animal models; disease models; laboratory animals; experimental infections; immunization; vaccination; vaccines; immune sensitization; Severe acute respiratory syndrome coronavirus 2; man; Escherichia coli; Betacoronavirus; Severe acute respiratory syndrome-related coronavirus; Coronavirinae; Coronaviridae; Nidovirales; positive-sense ssRNA Viruses; ssRNA Viruses; RNA Viruses; viruses; Homo; Hominidae; primates; mammals; vertebrates; Chordata; animals; eukaryotes; Escherichia; Enterobacteriaceae; Enterobacteriales; Gammaproteobacteria; Proteobacteria; Bacteria; prokaryotes; antigenicity; immunogens; SARS-CoV-2; viral infections; antigenic determinants; T cells; B cells; E. coli
Collection:
Bases de données des oragnisations internationales
Base de données:
CAB Abstracts
Type d'étude:
Études expérimentales
Les sujets:
Vaccins
langue:
Persan
Revue:
Koomesh
Année:
2022
Type de document:
Article
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