Robust and low-cost ELISA based on IgG-Fc tagged recombinant proteins to screen for anti-SARS-CoV-2 antibodies.
J Immunol Methods
; 495: 113082, 2021 08.
Article
in English
| MEDLINE | ID: covidwho-1243047
ABSTRACT
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 SARS-CoV-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.
Keywords
Full text:
Available
Collection:
International databases
Database:
MEDLINE
Main subject:
Recombinant Proteins
/
Immunoglobulin G
/
Immunoglobulin Fc Fragments
/
Enzyme-Linked Immunosorbent Assay
/
COVID-19 Serological Testing
/
SARS-CoV-2
/
COVID-19
Type of study:
Diagnostic study
Limits:
Animals
/
Humans
Language:
English
Journal:
J Immunol Methods
Year:
2021
Document Type:
Article
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