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. 2020 Sep;58(1):e108.
doi: 10.1002/cpmc.108.

An In Vitro Microneutralization Assay for SARS-CoV-2 Serology and Drug Screening

Fatima Amanat  1   2 , Kris M White  2   3 , Lisa Miorin  2   3 , Shirin Strohmeier  2 , Meagan McMahon  2 , Philip Meade  1   2 , Wen-Chun Liu  2   3 , Randy A Albrecht  2   3 , Viviana Simon  2   3   4 , Luis Martinez-Sobrido  5 , Thomas Moran  2 , Adolfo García-Sastre  2   3   4   6 , Florian Krammer  2
Affiliations

An In Vitro Microneutralization Assay for SARS-CoV-2 Serology and Drug Screening

Fatima Amanat et al. Curr Protoc Microbiol. 2020 Sep.

Abstract

The severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) emerged in the city of Wuhan, Hubei Province, China, in late 2019. Since then, the virus has spread globally and caused a pandemic. Assays that can measure the antiviral activity of antibodies or antiviral compounds are needed for SARS-CoV-2 vaccine and drug development. Here, we describe in detail a microneutralization assay, which can be used to assess in a quantitative manner if antibodies or drugs can block entry and/or replication of SARS-CoV-2 in vitro. © 2020 Wiley Periodicals LLC. Basic Protocol 1: Microneutralization assay to test inhibition of virus by antibodies (purified antibodies or serum/plasma) Basic Protocol 2: Screening of anti-SARS-CoV-2 compounds in vitro Support Protocol: SARS-CoV-2 propagation.

Keywords: COVID-19; COVID19; SARS-CoV-2; antivirals; medium-throughput screening; microneutralization; neutralization.

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Conflict of interest statement

Mount Sinai has licensed serological assays to commercial entities and has filed for patent protection for serological assays.

Figures

Figure 1
Figure 1
CPE induced by SARS‐CoV‐2 in Vero.E6 cells. The left panel shows healthy, uninfected Vero.E6 cells forming a monolayer. The right panel shows CPE after infection with SARS‐CoV‐2 (e.g., rounded up cells, halo around cells, etc.).
Figure 2
Figure 2
Plate layout for microneutralization assay. Indicated dilutions are suggestions for testing serum samples. Other dilution series or concentrations (for mAbs) might be used.
Figure 3
Figure 3
Analysis of data as reciprocal dilution of serum and percent inhibition of virus (ID50 of serum sample 1 is 1:298 and serum sample 2 is 1:1873).
Figure 4
Figure 4
Plate X and plate Y layout for the antiviral assay.
See this image and copyright information in PMC

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