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Review
. 2022 Jul 18;14(7):1560.
doi: 10.3390/v14071560.

Overview of Neutralization Assays and International Standard for Detecting SARS-CoV-2 Neutralizing Antibody

Kuan-Ting Liu  1   2 , Yi-Ju Han  1   2 , Guan-Hong Wu  1   2 , Kuan-Ying A Huang  1   3 , Peng-Nien Huang  1   3   4
Affiliations
Review

Overview of Neutralization Assays and International Standard for Detecting SARS-CoV-2 Neutralizing Antibody

Kuan-Ting Liu et al. Viruses. .

Abstract

We aimed to review the existing literature on the different types of neutralization assays and international standards for severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). We comprehensively summarized the serological assays for detecting neutralizing antibodies against SARS-CoV-2 and demonstrated the importance of an international standard for calibrating the measurement of neutralizing antibodies. Following the coronavirus disease outbreak in December 2019, there was an urgent demand to detect neutralizing antibodies in patients or vaccinated people to monitor disease outcomes and determine vaccine efficacy. Therefore, many approaches were developed to detect neutralizing antibodies against SARS-CoV-2, such as microneutralization assay, SARS-CoV-2 pseudotype virus assay, enzyme-linked immunosorbent assay (ELISA), and rapid lateral flow assay. Given the many types of serological assays for quantifying the neutralizing antibody titer, the comparison of different assay results is a challenge. In 2020, the World Health Organization proposed the first international standard as a common unit to define neutralizing antibody titer and antibody responses against SARS-CoV-2. These standards are useful for comparing the results of different assays and laboratories.

Keywords: international standard for SARS-CoV-2; neutralizing antibody detection; serological assays.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
The principle of neutralizing antibody detection using ELISA. Based on the competitive ELISA, one of the methods (A), the HRP-ACE2 competes with SARS-CoV-2 neutralizing antibody to bind to the antigen-like S1 or RBD. The light signal is strong when the level of the neutralizing antibody is low. In the other method (B), the neutralizing antibody is pre-incubated with antigen-like S1 or RBD and then added to the well that is coated with ACE2. After the wash step, the antigens not recognized by the neutralizing antibodies can bind to the well plate (ACE2), and a bright color can also be detected. ELISA, enzyme-linked immunosorbent assay; HRP-ACE2, horseradish peroxidase-angiotensin-converting enzyme 2; RBD, receptor binding domain; SARS-CoV-2, severe acute respiratory syndrome coronavirus 2; S1, spike protein 1.
Figure 2
Figure 2
The scheme of neutralizing antibody LFA. After the drop of serum sample is placed on the sample pad, the sample moves to the conjugate pad by capillary action. In this part, the AuNP-antigen can be recognized by the neutralizing antibody. Antibody detection, including IgM and IgG, occurs in the next part; the antibody and AuNP-antigen complex show a signal in the test region. AuNP, gold nanoparticle; Ig, immunoglobulin; LFA, lateral flow assay; SARS-CoV-2, severe acute respiratory syndrome corona virus 2.
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Grants and funding

This research received funding from the Chang Gung Medical Research Program (CORPD1J0061-63). This work was financially supported by the Research Center for Emerging Viral Infections from The Featured Areas Research Center Program within the framework of the Higher Education Sprout Project by the Ministry of Education (MOE) in Taiwan and the Ministry of Science and Technology (MOST), Taiwan (MOST 111-2634-F-182-001).