Rapid and Quantitative In Vitro Evaluation of SARS-CoV-2 Neutralizing Antibodies and Nanobodies
| Autor: | Weishu Wu, Xiaotian Tan, Jennifer Zupancic, John S. Schardt, Alec A. Desai, Matthew D. Smith, Jie Zhang, Liangzhi Xie, Maung Khaing Oo, Peter M. Tessier, Xudong Fan |
|---|---|
| Rok vydání: | 2022 |
| Předmět: | |
| Zdroj: | Anal Chem |
| ISSN: | 1520-6882 0003-2700 |
| DOI: | 10.1021/acs.analchem.2c00062 |
| Popis: | 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 hours, does not require BSL-2 facilities, and consumes only 8 μL of 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 IC(50) 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 rapid identification of an immune-evasive mutation of the SARS-CoV-2 spike protein utilizing a set of nanobodies with known binding epitopes. |
| Databáze: | OpenAIRE |
| Externí odkaz: |
|