Your browser doesn't support javascript.
Recombinant highly antigenic truncated fusion-based protein as a diagnostic antigen for anti-SARS-CoV-2 nucleocapsid antibody ELISA.
Salarifar, Abbasali; Khalili, Saeed; Rasaee, Mohammad Javad.
  • Salarifar A; Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran.
  • Khalili S; Department of Biology Sciences, Shahid Rajaee Teacher Training University, Tehran, Iran.
  • Rasaee MJ; Department of Clinical Biochemistry, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran. Electronic address: Rasaee_m@modares.ac.ir.
Protein Expr Purif ; 203: 106200, 2022 Nov 13.
Article in English | MEDLINE | ID: covidwho-2235165
ABSTRACT
Among the main structural protein of severe acute respiratory syndrome coronavirus-2 (SARS-CoV-2), nucleocapsid phosphoprotein (NP) exhibits high immunogenicity and is the most abundant viral protein produced and shed during infection. Detection of antibodies against NP may help assess the number of individuals exposed to SARS-COV-2 or vaccinated against it. Based on these findings and other structural and antigenic evaluations, we designed a recombinant truncated fusion NP-based protein for application in an immunoassay for detecting immunoglobulins in patients who have recovered from COVID-19. In this research, we aligned the NPs from SARS-CoV and SARS-CoV-2 and selected highly antigenic parts of the SARS-CoV-2 sequences based on in-silico studies. The protein was expressed under optimum conditions in the bacterial host BL21 and purified by nickel immobilized metal affinity chromatography. Moreover, the purity level was assessed by SDS-PAGE and Western blotting whereas the folding of the protein was evaluated by circular dichroism. Ultimately, we used the purified recombinant protein in ELISA development in which 42 samples from convalescent patients were compared with 20 samples of the past 2019 patients who had attended laboratories for various clinical check-ups. The sensitivity and specificity were determined as 71% and 90%, respectively, in the optimum cut-off point measured by the receiver operating characteristic curve.
Keywords

Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Experimental Studies / Prognostic study Topics: Vaccines Language: English Journal: Protein Expr Purif Journal subject: Molecular Biology Year: 2022 Document Type: Article Affiliation country: J.pep.2022.106200

Similar

MEDLINE

...
LILACS

LIS


Full text: Available Collection: International databases Database: MEDLINE Type of study: Diagnostic study / Experimental Studies / Prognostic study Topics: Vaccines Language: English Journal: Protein Expr Purif Journal subject: Molecular Biology Year: 2022 Document Type: Article Affiliation country: J.pep.2022.106200