Preprint Article Version 1 Preserved in Portico This version is not peer-reviewed

Application of an Rbd-Based Simple Immunoassay for Assessing Humoral Immunity against Emerging SARS-CoV-2 Virus Variants

Orsolya Mózner
ORCID logo ,
Judit Moldvay
ORCID logo ,
Kata Sára Szabó
ORCID logo ,
Dorottya Vaskó
,
Júlia Domján
,
Dorottya Ács
,
Zoltán Ligeti
,
Csaba Fehér
ORCID logo ,
Edit Hirsch
,
László Puskás
,
Cordula Stahl
,
Manfred Frey
ORCID logo ,
Balázs Sarkadi
* ORCID logo
Version 1 : Received: 29 October 2023 / Approved: 30 October 2023 / Online: 30 October 2023 (07:06:58 CET)

A peer-reviewed article of this Preprint also exists.

Mózner, O.; Moldvay, J.; Szabó, K.S.; Vaskó, D.; Domján, J.; Ács, D.; Ligeti, Z.; Fehér, C.; Hirsch, E.; Puskás, L.; Stahl, C.; Frey, M.; Sarkadi, B. Application of A Receptor-Binding-Domain-Based Simple Immunoassay for Assessing Humoral Immunity against Emerging SARS-CoV-2 Virus Variants. Biomedicines 2023, 11, 3193. Mózner, O.; Moldvay, J.; Szabó, K.S.; Vaskó, D.; Domján, J.; Ács, D.; Ligeti, Z.; Fehér, C.; Hirsch, E.; Puskás, L.; Stahl, C.; Frey, M.; Sarkadi, B. Application of A Receptor-Binding-Domain-Based Simple Immunoassay for Assessing Humoral Immunity against Emerging SARS-CoV-2 Virus Variants. Biomedicines 2023, 11, 3193.

Abstract

We have developed a simple, rapid, high-throughput RBD-based ELISA to assess the humoral immunity against emerging SARS-CoV-2 virus variants. The cDNAs of the his-tagged RBD proteins of the virus variants were stably engineered into HEK cells secreting the protein into the supernatant, and RBD purification was performed by Ni-chromatography and buffer exchange by membrane filtration. The simplified assay uses single dilutions of sera from finger-pricked native blood samples, purified RBD in 96-well plates, and a chromogenic dye for development. The results of this RBD-ELISA were confirmed to correlate with those of a commercial immunoassay measuring antibodies against the Wuhan strain, as well as direct virus neutralization assays assessing the cellular effects of the Wuhan and the Omicron (BA.5) variants. Here we document the applicability of this ELISA to assess the variant-specific humoral immunity in vaccinated and convalescent patients, as well as to follow the time course of selective vaccination response. This simple and rapid assay, easily modified to detect humoral immunity against emerging SARS-CoV-2 virus variants, may help to assess the level of antiviral protection after vaccination or infection.

Keywords

SARS-CoV-2 immunity; ELISA; Spike-RBD; SARS-CoV-2 variants; Omicron; virus neutralizing

Subject

Medicine and Pharmacology, Epidemiology and Infectious Diseases

Copyright: This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.

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