期刊
VIRUSES-BASEL
卷 12, 期 5, 页码 -出版社
MDPI
DOI: 10.3390/v12050513
关键词
SARS-CoV-2; COVID-19; coronavirus; neutralization assay; lentiviral pseudotype; Spike; cytoplasmic tail; ACE2; 293T-ACE2; luciferase; ALAYT
类别
资金
- NIAID of the NIH [R01AI141707, F30AI149928, HHSN272201700059C]
- National Institute of General Medical Sciences [R01GM120553]
- Pew Biomedical Scholars Award
- Investigators in the Pathogenesis of Infectious Disease Awards from the BurroughsWellcome Fund
- National Institutes for Drug Abuse (NIDA) Avenir New Innovator Award [DP2DA040254]
SARS-CoV-2 enters cells using its Spike protein, which is also the main target of neutralizing antibodies. Therefore, assays to measure how antibodies and sera affect Spike-mediated viral infection are important for studying immunity. Because SARS-CoV-2 is a biosafety-level-3 virus, one way to simplify such assays is to pseudotype biosafety-level-2 viral particles with Spike. Such pseudotyping has now been described for single-cycle lentiviral, retroviral, and vesicular stomatitis virus (VSV) particles, but the reagents and protocols are not widely available. Here, we detailed how to effectively pseudotype lentiviral particles with SARS-CoV-2 Spike and infect 293T cells engineered to express the SARS-CoV-2 receptor, ACE2. We also made all the key experimental reagents available in the BEI Resources repository of ATCC and the NIH. Furthermore, we demonstrated how these pseudotyped lentiviral particles could be used to measure the neutralizing activity of human sera or plasma against SARS-CoV-2 in convenient luciferase-based assays, thereby providing a valuable complement to ELISA-based methods that measure antibody binding rather than neutralization.
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