Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies

被引:8
作者
Frumence, Etienne [1 ,2 ,3 ]
Lebeau, Gregorie [1 ,2 ,3 ]
Viranaicken, Wildriss [4 ]
Dobi, Anthony [1 ,2 ,3 ]
Vagner, Damien [3 ,4 ]
Rakoto, Mahary Lalarizo [1 ,2 ,3 ,5 ]
Seteyen, Anne-Laure Sandenon [1 ,2 ,3 ]
Giry, Claude [6 ]
Septembre-Malaterre, Axelle [1 ,2 ,3 ]
Raffray, Loic [4 ,7 ]
Gasque, Philippe [1 ,2 ,3 ]
机构
[1] Univ Reunion, Unite Rech Pharmacoimmunol, F-97400 St Denis, France
[2] CHU Reunion, F-97400 St Denis, France
[3] CHU Reunion, Pole Biol, Lab Immunol Clin & Expt ZOI LIC, F-97400 St Denis, France
[4] Univ Reunion, CNRS 9192, INSERM 1187, UMR PIMIT Proc Infect Milieu Insulaire Trop, F-97400 St Denis, France
[5] Univ Antananarivo, Fac Med, Antananarivo, Madagascar
[6] CHU Reunion, Pole Biol, Lab Microbiol, CNR Associe Arbovirus, F-97440 St Denis, France
[7] Ctr Hosp Univ Reunion, Serv Med Interne, Site Felix Guyon,CS11021, F-97400 St Denis, France
关键词
CHO cells; COVID-19; ELISA; Recombinant protein; SARS-CoV-2; Serological assay; NUCLEOCAPSID PROTEIN; CORONAVIRUS;
D O I
10.1016/j.jim.2021.113082
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
The development of new diagnostic assays become a priority for managing COVID-19. To this aim, we presented here an in-house ELISA based on the production of two major recombinant and high-quality antigens from SARSCoV-2. Full-length N and S-RBD fragment proteins fused to mouse IgG2a-Fc were produced in the CHO cell line. Secreted recombinant proteins were easily purified with standard Protein A chromatography and were used in an in-house ELISA to detect anti-N and anti-RBD IgGs in the plasma of COVID-19 RTPCR-positive patients. High reactivity against recombinant antigens was readily detected in all positive plasma samples, whereas no recognition was observed with control healthy subject's plasmas. Remarkably, unpurified recombinant N protein obtained from cell culture supernatant was also suitable for the monitoring by ELISA of IgG levels in positive patients. This work provides an early prospection for low price but high-quality serological kit development.
引用
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页数:6
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共 17 条
[1]   Human secretory signal peptide description by hidden Markov model and generation of a strong artificial signal peptide for secreted protein expression [J].
Barash, S ;
Wang, W ;
Shi, YG .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2002, 294 (04) :835-842
[2]   Sensitivity in Detection of Antibodies to Nucleocapsid and Spike Proteins of Severe Acute Respiratory Syndrome Coronavirus 2 in Patients With Coronavirus Disease 2019 [J].
Burbelo, Peter D. ;
Riedo, Francis X. ;
Morishima, Chihiro ;
Rawlings, Stephen ;
Smith, Davey ;
Das, Sanchita ;
Strich, Jeffrey R. ;
Chertow, Daniel S. ;
Davey, Richard T., Jr. ;
Cohen, Jeffrey, I .
JOURNAL OF INFECTIOUS DISEASES, 2020, 222 (02) :206-213
[3]   Fc-fusion proteins: new developments and future perspectives [J].
Czajkowsky, Daniel M. ;
Hu, Jun ;
Shao, Zhifeng ;
Pleass, Richard J. .
EMBO MOLECULAR MEDICINE, 2012, 4 (10) :1015-1028
[4]   Cell type-specific cleavage of nucleocapsid protein by effector caspases during SARS coronavirus infection [J].
Diemer, Claudia ;
Schneider, Martha ;
Seebach, Judith ;
Quaas, Janine ;
Froesner, Gert ;
Schaetzl, Hermann M. ;
Gilch, Sabine .
JOURNAL OF MOLECULAR BIOLOGY, 2008, 376 (01) :23-34
[5]   A Chimeric Zika Virus between Viral Strains MR766 and BeH819015 Highlights a Role for E-glycan Loop in Antibody-mediated Virus Neutralization [J].
Frumence, Etienne ;
Viranaicken, Wildriss ;
Bos, Sandra ;
Alvarez-Martinez, Maria-Teresa ;
Roche, Marjolaine ;
Arnaud, Jacques-Damien ;
Gadea, Gilles ;
Despres, Philippe .
VACCINES, 2019, 7 (02)
[6]   USE OF THE HUMAN ELONGATION FACTOR-1-ALPHA PROMOTER AS A VERSATILE AND EFFICIENT EXPRESSION SYSTEM [J].
KIM, DW ;
UETSUKI, T ;
KAZIRO, Y ;
YAMAGUCHI, N ;
SUGANO, S .
GENE, 1990, 91 (02) :217-223
[7]   Deciphering SARS-CoV-2 Virologic and Immunologic Features [J].
Lebeau, Gregorie ;
Vagner, Damien ;
Frumence, Etienne ;
Ah-Pine, Franck ;
Guillot, Xavier ;
Nobecourt, Estelle ;
Raffray, Loic ;
Gasque, Philippe .
INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES, 2020, 21 (16) :1-40
[8]   Antibody response of patients with severe acute respiratory syndrome (SARS) targets the viral nucleocapsid [J].
Leung, DTM ;
Tam, FCH ;
Ma, CH ;
Chan, PKS ;
Cheung, JLK ;
Niu, HT ;
Tam, JSL ;
Lim, PL .
JOURNAL OF INFECTIOUS DISEASES, 2004, 190 (02) :379-386
[9]  
Long Quan-Xin, 2020, NAT MED, V26, P845, DOI DOI 10.1186/s12888-018-1699-6
[10]   SARS coronavirus: Unusual lability of the nucleocapsid protein [J].
Mark, John ;
Li, Xuguang ;
Cyr, Terry ;
Fournier, Sylvie ;
Jaentschke, Lozena ;
Hefford, Mary Alice .
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS, 2008, 377 (02) :429-433