Protocol and Reagents for Pseudotyping Lentiviral Particles with SARS-CoV-2 Spike Protein for Neutralization Assays

• Published in: Viruses Vol. 12; no. 5; p. 513
• Main Authors: Crawford, Katharine H. D., Eguia, Rachel, Dingens, Adam S., Loes, Andrea N., Malone, Keara D., Wolf, Caitlin R., Chu, Helen Y., Tortorici, M. Alejandra, Veesler, David, Murphy, Michael, Pettie, Deleah, King, Neil P., Balazs, Alejandro B., Bloom, Jesse D.
• Format: Journal Article
• Language: English
• Published: Switzerland MDPI AG 06.05.2020; MDPI
• Subjects: Angiotensin-Converting Enzyme 2; Antibodies, Neutralizing; Antibodies, Neutralizing - immunology; Antibodies, Viral; Antibodies, Viral - blood; Antibodies, Viral - immunology; Biochemical assays; Containment of Biohazards; coronavirus; Coronaviruses; COVID-19; Diagnosis; Health aspects; HEK293 Cells; Humans; lentiviral pseudotype; Lentivirus; Life Sciences; Methods; Microbiology and Parasitology; neutralization assay; Neutralization Tests; Neutralization Tests - methods; Peptidyl-Dipeptidase A; Peptidyl-Dipeptidase A - metabolism; Plasma; Plasma - immunology; Protocol; SARS-CoV-2; Spike; Spike Glycoprotein, Coronavirus; Spike Glycoprotein, Coronavirus - analysis; Virology; United States; COVID-19; Spike; ACE2; SARS-CoV-2; cytoplasmic tail; ALAYT; lentiviral pseudotype; luciferase; coronavirus; neutralization assay; 293T-ACE2

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 simpl...