Pseudovirus Assay
Pseudovirus Assay - Related Videos
02:43
Luciferase-Based Pseudovirus Neutralization Assay
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2026
Source: Chang, X. et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021)The video demonstrates the neutralization of influenza pseudoviruses by antibodies that block the HA protein from binding to host cells. A luciferase reporter system reveals the reduction in viral entry with increasing antibody concentrations, showing the effectiveness of antibody-mediated neutralization.
02:01
Assessing Pseudovirus Infection Using Luciferase Reporter Assay
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021).This video demonstrates a luciferase-based assay to quantify pseudovirus infection in epithelial cells. The pseudovirus encodes the luciferase gene, which is expressed in infected cells. After washing, and cells are incubated with lysis buffer at an ultra-low temperature, and then thawed to induce lysis.
03:25
Quantification of Pseudovirus Particles by Plaque Assay
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2026
Source: Jamieson, T. R., et al., Detection of SARS-CoV-2 Neutralizing Antibodies using High-Throughput Fluorescent Imaging of Pseudovirus Infection. J. Vis. Exp. (2021)This video describes the virus plaque assay using coronavirus pseudovirus particles. The diluted pseudovirus is added to a monolayer of mammalian cells, followed by incubation with a methylcellulose overlay. Plaques formed by infected cells are counted visually to determine the infectious virus titer.
03:09
Determining Optimal Target Cell Density Using a Luciferase-Based Pseudovirus Infection Assay
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021)This video demonstrates a luciferase-based pseudovirus titration assay to determine the optimal target cell density for maximum infection. By comparing luminescence across cell densities, the method ensures high assay sensitivity and reproducibility for downstream applications.
02:22
Production of Pseudovirus Using Plasmid Transfection in Human Cells
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021)This video demonstrates the production of influenza pseudoviruses in human cells using calcium phosphate-mediated plasmid transfection, enabling safe generation of single-cycle viral particles for downstream applications such as antibody neutralization assays under biosafety level 2 conditions.
FAQs
The assay changes the surface protein displayed on the engineered particle or examines entry into susceptible cells with different receptor properties. Reporter expression then indicates whether attachment and entry occurred. This makes the system useful for linking a pathogen surface protein to receptor usage and for studying how entry-related mechanisms contribute to infection.
Reporter expression provides a quantifiable readout of successful pseudovirus entry into susceptible cells. A stronger signal indicates greater entry efficiency under the tested conditions, whereas a reduced signal in the presence of antibodies, sera, or an entry inhibitor indicates that the tested material interfered with entry. The result therefore connects molecular activity with measurable cell entry.
Replication-defective particles allow researchers to examine virus entry or neutralization without producing a complete infectious virus. This distinction can improve biosafety and supports standardized comparisons across experiments. The format focuses measurements on the entry step, making it useful for analyzing surface-protein function, receptor usage, and factors that block initial infection-related events.
A typical workflow selects a pathogen surface protein, incorporates it onto engineered particles, and includes a reporter gene for detection. The particles are then applied to susceptible cells, where entry can produce reporter expression. Researchers measure the resulting signal and compare it with conditions containing antibodies, sera, or entry inhibitors to assess entry or neutralization.
Researchers expose the pseudovirus system to vaccine-elicited antibodies or sera and assess whether reporter expression decreases after the particles contact susceptible cells. A lower signal reflects reduced entry and therefore supports analysis of neutralizing activity. Because the assay is adaptable and quantifiable, it can help compare immune responses across sera or antibody preparations.
The method is useful when investigators need to compare receptor usage, antibody neutralization, vaccine-elicited immune responses, sera, or entry inhibitors. Its reporter-based output provides a measurable indicator of entry-related activity, while its replication-defective design supports work focused on infection mechanisms and immune protection without requiring production of a complete infectious virus.