Hybrid alphavirus-SARS-CoV-2 particle and methodology of making and using same
Inventors
Assignees
George Mason University • George Mason Research Foundation Inc
Publication Number
US-11473064-B2
Publication Date
2022-10-18
Expiration Date
2041-10-19
Interested in licensing this patent?
MTEC can help explore whether this patent might be available for licensing for your application.
Abstract
Timely development of vaccines and antiviral drugs is critical to control the COVID-19 pandemic. Current methods for quantifying vaccine-induced neutralizing antibodies involve the use of pseudoviruses, such as the SARS-CoV-2 spike protein (S) pseudotyped lentivirus. However, these pseudoviruses contain structural proteins foreign to SARS-CoV-2, and require days to infect and express reporter genes. Here, the present application discloses composition and methodology for making and using a new hybrid alphavirus-SARS-CoV-2 (Ha-CoV-2) particle for rapid and accurate quantification of neutralization antibodies and viral variants.
Core Innovation
The invention discloses a novel hybrid alphavirus-SARS-CoV-2 (Ha-CoV-2) particle designed for rapid and accurate quantification of neutralizing antibodies and viral variants. This hybrid particle is a non-replicating SARS-CoV-2 virus-like particle (VLP) that incorporates one or more authentic virus structural proteins (S, M, N, and E) from SARS-CoV-2, combined with an RNA genome derived from an alphavirus-based vector. The alphavirus vector facilitates rapid and robust expression of reporter genes such as GFP or luciferase in target cells within hours of viral entry.
The hybrid particle platforms address significant limitations of conventional methods for quantifying vaccine-induced neutralizing antibodies, which typically employ pseudoviruses like lentiviruses or vesicular stomatitis virus containing only the S protein of SARS-CoV-2. These conventional pseudoviruses have structural proteins foreign to SARS-CoV-2, take days to generate reporter signals, and can produce false positives or limited sensitivity, particularly for cells expressing native levels of ACE2.
By structurally mimicking SARS-CoV-2 more closely and enabling rapid reporter expression, Ha-CoV-2 serves as a robust platform for fast and sensitive quantification of neutralizing antibodies, viral variant infectivity, antiviral drug screening, and evaluation of immune response. The methodology supports assembling variant-specific particles to monitor the impact of mutations and enables applications such as rapid screening, large-scale diagnostics, and vaccine-related studies.
Claims Coverage
There are three primary inventive features in the independent claims of this patent, focusing on hybrid particle composition, methods of production and use, and composition as a vaccine or immune-stimulating agent.
Hybrid alphavirus-SARS-CoV-2 particle composition
A hybrid particle comprising: - An RNA genome derived from an alphavirus, provided by a first plasmid. - At least one SARS-CoV-2 structural protein (S, M, E, or N), provided by a second plasmid differing from the first plasmid. - The hybrid particle is a hybrid pseudovirus assembled and produced in a producer cell by co-transfection with these plasmids.
Method for making and using a hybrid particle with rapid reporter expression
A method comprising: 1. Co-transfecting a first vector encoding at least one SARS-CoV-2 structural protein and a second vector comprising an RNA genome derived from an alphavirus. 2. The alphavirus RNA genome comprises: - A DNA promoter at the 5′ end - 5′ untranslated region - Open reading frame coding a nonstructural protein from an alphavirus - Reporter gene - SARS-CoV-2 packaging signal sequence - Viral subgenomic RNA promoter - Gene of interest - 3′ untranslated region - PolyA tail 3. Generating the hybrid particle as described.
Hybrid particle as a vaccine or immune response-initiating composition
A composition comprising the hybrid alphavirus-SARS-CoV-2 particle, where: - The particle contains at least one or specifically four SARS-CoV-2 structural proteins (S, E, M, and N). - The composition can function as a vaccine or as a particle that can initiate an immune response.
The inventive features claim a hybrid particle composed of SARS-CoV-2 structural proteins and alphavirus-derived genome, methods for assembling such particles for rapid detection and quantification applications, and compositions for use as vaccines or immune-stimulating agents.
Stated Advantages
The hybrid VLPs enable rapid and robust expression of reporter genes within hours of viral entry, allowing rapid screening and quantification of neutralizing antibodies and antiviral drugs.
Ha-CoV-2 particles closely resemble authentic SARS-CoV-2, overcoming limitations of pseudoviruses that contain structural proteins foreign to SARS-CoV-2.
The platform provides a tool for rapid evaluation of SARS-CoV-2 variants and assessment of impacts on vaccine effectiveness and antibody neutralization.
The particles can be produced cheaply and at large scale, supporting commercial and clinical applications.
Ha-CoV-2 particles can be used for high-throughput drug screening, vaccine discovery, and modeling SARS-CoV-2 infection pathways.
The system can facilitate personalized medicine approaches by determining antibody presence against panel of SARS-CoV-2 variants.
Documented Applications
Rapid quantification and screening of neutralizing antibodies against SARS-CoV-2 and its variants.
Screening of SARS-CoV-2 entry inhibitors, antiviral drugs, and evaluation of their inhibitory activities.
Modeling SARS-CoV-2 infection pathways for research in drug and vaccine development.
Measuring subject antibody responses to SARS-CoV-2 variants for purposes such as personalized medicine or travel risk assessment.
Use as a vaccine or as a particle to initiate an immune response.
Interested in licensing this patent?