4.8 Article

Rapid and Quantitative In Vitro Evaluation of SARS-CoV-2 Neutralizing Antibodies and Nanobodies

Journal

ANALYTICAL CHEMISTRY
Volume 94, Issue 10, Pages 4504-4512

Publisher

AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.2c00062

Keywords

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Funding

  1. National Science Foundation [ECCS 2029484, CBET 1159943, 1605266, 1813963]
  2. National Institutes of Health [RF1AG059723, R35GM136300, F32 GM137513]
  3. Albert M. Mattocks Chair
  4. Biointerfaces Institute
  5. Div Of Chem, Bioeng, Env, & Transp Sys
  6. Directorate For Engineering [1813963] Funding Source: National Science Foundation

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In this study, a rapid and quantitative assay was developed to evaluate the neutralizing efficacy of antibodies and nanobodies against COVID-19. The assay showed comparable results to traditional virus neutralization tests and demonstrated its ability to assess the efficacy against different SARS-CoV-2 variants.
Neutralizing monoclonal antibodies and nanobodies have shown promising results as potential therapeutic agents for COVID-19. Identifying such antibodies and nanobodies requires evaluating the neutralization activity of a large number of lead molecules via biological assays, such as the virus neutralization test (VNT). These assays are typically time-consuming and demanding on-lab facilities. Here, we present a rapid and quantitative assay that evaluates the neutralizing efficacy of an antibody or nanobody within 1.5 h, does not require BSL-2 facilities, and consumes only 8 mu L of a low concentration (ng/mL) sample for each assay run. We tested the human angiotensin-converting enzyme 2 (ACE2) binding inhibition efficacy of seven antibodies and eight nanobodies and verified that the IC50 values of our assay are comparable with those from SARS-CoV-2 pseudovirus neutralization tests. We also found that our assay could evaluate the neutralizing efficacy against three widespread SARS-CoV-2 variants. We observed increased affinity of these variants for ACE2, including the beta and gamma variants. Finally, we demonstrated that our assay enables the rapid identification of an immune-evasive mutation of the SARS-CoV-2 spike protein, utilizing a set of nanobodies with known binding epitopes.

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