Abstract
<p>This protocol describes a 96-well plate-based pseudovirus neutralization assay using VSV-ΔG or lentivirus pseudotyped with a target viral glycoprotein and a luciferase reporter gene. Neutralizing antibodies block pseudovirus entry into receptor-expressing target cells, reducing luciferase signal in a dose-dependent manner. The assay generates IC₅₀ values via 4-parameter logistic (4PL) regression and is the standard format for vaccine-induced neutralizing antibody measurement and therapeutic antibody screening without BSL-3 containment. A successful assay produces a sigmoidal dose-response curve, with IC₅₀ values typically ranging from 0.01–1 μg/mL for potent neutralizing antibodies.This protocol describes a 96-well plate-based pseudovirus neutralization assay using VSV-ΔG or lentivirus pseudotyped with a target viral glycoprotein and a luciferase reporter gene. Neutralizing antibodies block pseudovirus entry into receptor-expressing target cells, reducing luciferase signal in a dose-dependent manner. The assay generates IC₅₀ values via 4-parameter logistic (4PL) regression and is the standard format for vaccine-induced neutralizing antibody measurement and therapeutic antibody screening without BSL-3 containment. A successful assay produces a sigmoidal dose-response curve, with IC₅₀ values typically ranging from 0.01–1 μg/mL for potent neutralizing antibodies.</p>