ArticleMedComm2024
Optimization and validation of a virus-like particle pseudotyped virus neutralization assay for SARS-CoV-2.
Article in MedComm, 2024. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 10 papers.
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Who cites it
10 citing papers in PubMed.
- Comparative analysis of neutralization assays performed using live SARS-CoV-2 virus and pseudovirus to assess immunogenicity of a bivalent SARS-CoV-2 protein vaccine in humans.Frontiers in immunology · 2025Trial
- Article
- Measuring Humoral Immune Responses to SARS-CoV-2: A Comprehensive Review of Serological Assays.Vaccines · 2026Review
- Application of Orthoflavivirus Pseudovirus Technology in Antiviral Research.International journal of molecular sciences · 2026Review
- A multi-glycoprotein synergized recombinant virus system resolved the research dilemma of Epstein-Barr virus entry and facilitated the development of entry-specific drugs or vaccines.Current research in microbial sciences · 2026Article
- Neutralization of Crimean-Congo hemorrhagic fever pseudotyped virions with heavy chain antibodies.Virologica Sinica · 2025Article
- Pseudovirus-Based Neutralization Assays as Customizable and Scalable Tools for Serological Surveillance and Immune Profiling.Pathogens (Basel, Switzerland) · 2025Article
- Evaluation of the Dual Antiviral and Immunomodulatory Effects ofVeterinary sciences · 2025Article
- The Compensatory Effect of S375F on S371F Is Vital for Maintaining the Infectivity of SARS-CoV-2 Omicron Variants.Journal of medical virology · 2025Article
- Optimization and validation of a virus-like particle pseudotyped virus neutralization assay for SARS-CoV-2.MedComm · 2024Article
Corrections and comments
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Authors and funding
13 authors.
Funding
No grant is acknowledged in the PubMed record.
Abstract
Spike-protein-based pseudotyped viruses were used to evaluate vaccines during the COVID-19 pandemic. However, they cannot be used to evaluate the envelope (E), membrane (M), and nucleocapsid (N) proteins. The first generation of virus-like particle (VLP) pseudotyped viruses contains these four structural proteins, but their titers for wild-type severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) are relatively low, even lower for the omicron variant, rendering them unsuitable for neutralizing antibody detection. By optimizing the spike glycoprotein signal peptide, substituting the complexed M and E proteins with SARS-COV-1, optimizing the N protein with specific mutations (P199L, S202R, and R203M), and truncating the packaging signal, PS9, we increased the titer of the wild-type VLP pseudotyped virus over 100-fold, and successfully packaged the omicron VLP pseudotyped virus. The SARS-CoV-2 VLP pseudotyped viruses maintained stable titers, even through 10 freeze-thaw cycles. The key neutralization assay parameters were optimized, including cell type, cell number, and viral inoculum. The assay demonstrated minimal variation in both intra- and interassay results, at 11.5% and 11.1%, respectively. The correlation between the VLP pseudotyped virus and the authentic virus was strong (
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