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Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection
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SARS-CoV-2 Antibody Neutralization Assay Platforms Based on Epitopes Sources: Live Virus, Pseudovirus, and
Endah Puji Septisetyani1, Pekik Wiji Prasetyaningrum1, Khairul Anam1
1Research Center for Biotechnology, National Research and Innovation Agency, Bogor, West Java, Indonesia.
Abstract:
The high virulent severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) virus that emerged in China at the end of 2019 has generated novel coronavirus disease, coronavirus disease 2019 (COVID-19), causing a pandemic worldwide. Every country has made great efforts to struggle against SARS-CoV-2 infection, including massive vaccination, immunological patients' surveillance, and the utilization of convalescence plasma for COVID-19 therapy. These efforts are associated with the attempts to increase the titers of SARS-CoV-2 neutralizing Abs (nAbs) generated either after infection or vaccination that represent the body's immune status. As there is no standard therapy for COVID-19 yet, virus eradication will mainly depend on these nAbs contents in the body. Therefore, serological nAbs neutralization assays become a requirement for researchers and clinicians to measure nAbs titers. Different platforms have been developed to evaluate nAbs titers utilizing various epitopes sources, including neutralization assays based on the live virus, pseudovirus, and neutralization assays utilizing recombinant SARS-CoV-2 S glycoprotein receptor binding site, receptor-binding domain. As a standard neutralization assay, the plaque reduction neutralization test (PRNT) requires isolation and propagation of live pathogenic SARS-CoV-2 virus conducted in a BSL-3 containment. Hence, other surrogate neutralization assays relevant to the PRNT play important alternatives that offer better safety besides facilitating high throughput analyses. This review discusses the current neutralization assay platforms used to evaluate nAbs, their techniques, advantages, and limitations.
Insights
Measuring neutralizing antibodies (nAbs) is crucial for understanding COVID-19 immunity. This review details various SARS-CoV-2 neutralization assay platforms, their pros, and cons for researchers and clinicians.
Area of Science:
- Virology
- Immunology
- Infectious Diseases
Background:
- Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) caused a global pandemic, leading to coronavirus disease 2019 (COVID-19).
- Efforts to combat the pandemic include vaccination and therapies, focusing on increasing SARS-CoV-2 neutralizing antibodies (nAbs) for immune status assessment.
- nAbs are critical for virus eradication, necessitating reliable methods to measure their titers.
Purpose of the Study:
- To review current SARS-CoV-2 neutralization assay platforms for evaluating nAbs.
- To discuss the techniques, advantages, and limitations of various assay platforms.
Main Methods:
- Review of existing literature on SARS-CoV-2 neutralization assays.
- Comparison of different platforms including live virus, pseudovirus, and recombinant antigen-based assays.
- Discussion of the plaque reduction neutralization test (PRNT) and its surrogate alternatives.
Main Results:
- Multiple platforms exist to evaluate nAbs, utilizing diverse epitope sources.
- The plaque reduction neutralization test (PRNT) is a standard but requires BSL-3 containment.
- Surrogate neutralization assays offer safer, high-throughput alternatives to PRNT.
Conclusions:
- Accurate measurement of nAbs is essential for managing COVID-19.
- A variety of neutralization assays are available, each with specific applications.
- Surrogate assays provide practical advantages for widespread use in research and clinical settings.
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