tetano
Editor, Senior Moderator
J Med Virol
. 2020 Sep 23.
doi: 10.1002/jmv.26541. Online ahead of print.
N-terminally truncated nucleocapsid protein of SARS-CoV-2 as a better serological marker than whole nucleocapsid protein in evaluating the immunogenicity of inactive SARS-CoV-2
Lei Yue[SUP] 1 [/SUP], Han Cao[SUP] 1 [/SUP], Tianhong Xie[SUP] 1 [/SUP], Runxiang Long[SUP] 1 [/SUP], Hua Li[SUP] 1 [/SUP], Ting Yang[SUP] 1 [/SUP], Min Yan[SUP] 2 [/SUP], Zhongping Xie[SUP] 1 [/SUP]
Affiliations
Abstract
The coronavirus disease 2019 pandemic caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) had led to a serious public health crisis, and no specific treatments or vaccines are available yet. A nucleocapsid protein (NP)-based enzyme-linked immunosorbent assay (ELISA) detection method is not only important in disease diagnosis, but is required for the evaluation of vaccine efficacy during the development of an inactivated SARS-CoV-2 vaccine. In this study, we expressed both the NP and N-terminally truncated NP (ΔN-NP) of SARS-CoV-2 in an Escherichia coli expression system, and described the purification of the soluble recombinant NP and ΔN-NP in details. The identities of the NP and ΔN-NP were confirmed with mass spectrometry. We then used IgG detection ELISAs to compare the sensitivity of NP and ΔN-NP in detecting anti-SARS-CoV-2 antibodies. ΔN-NP showed greater sensitivity than NP in the analysis of serially diluted sera from mice and rabbits vaccinated with inactive SARS-CoV-2 and in human sera diluted 1:400. ΔN-NP showed a positive detection rate similar to that of the SARS-CoV-2 S protein in human sera. We conclude that ΔN-NP is a better serological marker than NP for evaluating the immunogenicity of inactivated SARS-CoV-2. This article is protected by copyright. All rights reserved.
Keywords: N-terminally truncated NP; Protein expression and purification; SARS-CoV-2; sensitivity.
. 2020 Sep 23.
doi: 10.1002/jmv.26541. Online ahead of print.
N-terminally truncated nucleocapsid protein of SARS-CoV-2 as a better serological marker than whole nucleocapsid protein in evaluating the immunogenicity of inactive SARS-CoV-2
Lei Yue[SUP] 1 [/SUP], Han Cao[SUP] 1 [/SUP], Tianhong Xie[SUP] 1 [/SUP], Runxiang Long[SUP] 1 [/SUP], Hua Li[SUP] 1 [/SUP], Ting Yang[SUP] 1 [/SUP], Min Yan[SUP] 2 [/SUP], Zhongping Xie[SUP] 1 [/SUP]
Affiliations
- PMID: 32965677
- DOI: 10.1002/jmv.26541
Abstract
The coronavirus disease 2019 pandemic caused by severe acute respiratory syndrome-coronavirus 2 (SARS-CoV-2) had led to a serious public health crisis, and no specific treatments or vaccines are available yet. A nucleocapsid protein (NP)-based enzyme-linked immunosorbent assay (ELISA) detection method is not only important in disease diagnosis, but is required for the evaluation of vaccine efficacy during the development of an inactivated SARS-CoV-2 vaccine. In this study, we expressed both the NP and N-terminally truncated NP (ΔN-NP) of SARS-CoV-2 in an Escherichia coli expression system, and described the purification of the soluble recombinant NP and ΔN-NP in details. The identities of the NP and ΔN-NP were confirmed with mass spectrometry. We then used IgG detection ELISAs to compare the sensitivity of NP and ΔN-NP in detecting anti-SARS-CoV-2 antibodies. ΔN-NP showed greater sensitivity than NP in the analysis of serially diluted sera from mice and rabbits vaccinated with inactive SARS-CoV-2 and in human sera diluted 1:400. ΔN-NP showed a positive detection rate similar to that of the SARS-CoV-2 S protein in human sera. We conclude that ΔN-NP is a better serological marker than NP for evaluating the immunogenicity of inactivated SARS-CoV-2. This article is protected by copyright. All rights reserved.
Keywords: N-terminally truncated NP; Protein expression and purification; SARS-CoV-2; sensitivity.