02178nas a2200217   4500008004100000245011100041210006900152260000800221300001200229490000800241520143400249100001801683700001501701700002001716700001801736700002101754700001701775700002001792700002001812856012801832       2022                        eng d00aDimerization of SARS-CoV-2 nucleocapsid protein affects sensitivity of ELISA based diagnostics of COVID-190 aDimerization of SARSCoV2 nucleocapsid protein affects sensitivit  cMAR  a428-4370 v2003 a<p>Nucleocapsid protein (N protein) is the primary antigen of the virus for development of sensitive diagnostic assays of COVID-19. In this paper, we demonstrate the significant impact of dimerization of the severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) N-protein on sensitivity of enzyme-linked immunosorbent assay (ELISA) based diagnostics. The expressed purified protein from E. coli is composed of dimeric and monomeric forms, which have been further characterized using biophysical and immunological techniques. Indirect ELISA indicated elevated susceptibility of the dimeric form of the nucleocapsid protein for identification of protein-specific monoclonal antibody as compared to the monomeric form. This finding also confirmed with the modelled structure of monomeric and dimeric nucleocapsid protein via HHPred software and its solvent accessible surface area, which indicates higher stability and antigenicity of the dimeric type as compared to the monomeric form. The sensitivity and specificity of the ELISA at 95% CI are 99.0% (94.5-99.9) and 95.0% (83.0-99.4), respectively, for the highest purified dimeric form of the N protein. As a result, using the highest purified dimeric form will improve the sensitivity of the current nucleocapsid-dependent ELISA for COVID-19 diagnosis, and manufacturers should monitor and maintain the monomer-dimer composition for accurate and robust diagnostics.</p>
1 aKhan, Wajihul1 aKhan, Nida1 aMishra, Avinash1 aGupta, Surbhi1 aBansode, Vikrant1 aMehta, Deepa1 aBhambure, Rahul1 aRathore, Anurag  uhttps://library.ncl.res.in/content/dimerization-sars-cov-2-nucleocapsid-protein-affects-sensitivity-elisa-based-diagnostics