cf96f2c1a9c66d3ee9b47edd50196030 pmc_11359663.pdf b80eb695dc9bf97757ca06b45576fe1c65c21cad pmc_11359663.pdf eddab0cc0520b0ced03d237dbce79cd5c9dd36a6d31f0914ef021fedeaccbd4b pmc_11359663.pdf Title: Gold Nanoparticle Virus-like Particles Presenting SARS-CoV-2 Spike Protein: Synthesis, Biophysical Properties and Immunogenicity in BALB/c Mice Subject: Gold nanoparticles (AuNPs) decorated with antigens have recently emerged as promising tools for vaccine development due to their innate ability to provide stability to antigens and modulate immune responses. In this study, we have engineered deactivated virus-like particles (VLPs) by precisely functionalizing gold cores with coronas comprising the full SARS-CoV-2 spike protein (S). Using BALB/c mice as a model, we investigated the immunogenicity of these S-AuNPs-VLPs. Our results demonstrate that S-AuNPs-VLPs consistently enhanced antigen-specific antibody responses compared to the S protein free in solution. This enhancement included higher binding antibody titers, higher neutralizing capacity of antibodies, and stronger T-cell responses. Compared to the mRNA COVID-19 vaccine, where the S protein is synthesized in situ, S-AuNPs-VLPs induced comparable binding and neutralizing antibody responses, but substantially superior T-cell responses. In conclusion, our study highlights the potential of conjugated AuNPs as an effective antigen-delivery system for protein-based vaccines targeting a broad spectrum of infectious diseases and other emergent viruses. Keywords: virus-like particles (VLPs); gold nanoparticles; spike protein SARS-CoV-2; SARS-CoV-2vaccine Author: Vivian A. Salazar, Joan Comenge, Rosa Suárez-López, Judith A. Burger, Rogier W. Sanders, Neus G. Bastús, Carlos Jaime, Joan Joseph-Munne and Victor Puntes Creator: LaTeX with hyperref Producer: pdfTeX-1.40.25 CreationDate: Tue Jul 23 11:10:25 2024 CEST ModDate: Tue Jul 23 11:16:20 2024 CEST Custom Metadata: no Metadata Stream: no Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 18 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 11818737 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- FLAGDG+VnURWPalladioL-Bold Type 1 Custom yes yes yes 10 0 RIHMCA+URWPalladioL-Bold Type 1 Custom yes yes yes 16 0 OZQASM+URWPalladioL-Roma Type 1 Custom yes yes yes 22 0 ORJENF+URWPalladioL-Ital Type 1 Custom yes yes yes 27 0 IYDCBB+VnURWPalladioL Type 1 Custom yes yes yes 32 0 QMDDXF+CMSY10 Type 1 Builtin yes yes yes 65 0 WPPLLU+EURM10 Type 1 Builtin yes yes yes 70 0 MPUDFL+PazoMath Type 1 Builtin yes yes yes 81 0 JHYTSG+CMR10 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RepresentationInformation: pmc_11359663.pdf ReportingModule: PDF-hul, Rel. 1.12.4 (2023-03-16) LastModified: 2024-09-18 02:30:40 CEST Size: 11818737 Format: PDF Version: 1.7 Status: Well-Formed and valid SignatureMatches: PDF-hul MIMEtype: application/pdf PDFMetadata: Objects: 491 FreeObjects: 1 IncrementalUpdates: 0 DocumentCatalog: PageLayout: SinglePage PageMode: UseNone Outlines: Item: Title: Introduction Destination: section.1 Item: Title: Materials and Methods Destination: section.2 Children: Item: Title: Synthesis of AuNPs Destination: subsection.2.1 Item: Title: Synthesis of S-AuNPs-VLPs Destination: subsection.2.2 Item: Title: Synthesis of Murine Albumin VLPs Destination: subsection.2.3 Item: Title: UV-Vis Characterization Destination: subsection.2.4 Item: Title: Dynamic Light Scattering (DLS) and Zeta Potential Analysis Destination: subsection.2.5 Item: Title: Sodium Cyanide Assay Destination: subsection.2.6 Item: Title: NaCl Stability Destination: subsection.2.7 Item: Title: Molecular Dynamics Destination: subsection.2.8 Item: Title: Bioconjugate Analysis by STEM-SEM Destination: subsection.2.9 Item: Title: Immunization of Mice, Collection of Sera, and Isolation of Splenocytes Destination: subsection.2.10 Item: Title: Enzyme-Linked Immunosorbent Assay Destination: subsection.2.11 Item: Title: Determination of Neutralizing Antibodies Destination: subsection.2.12 Item: Title: Antigen-Specific IFN Gamma Production Analysis Destination: subsection.2.13 Item: Title: Statistical Analysis Destination: subsection.2.14 Item: Title: Ethics Statements Destination: subsection.2.15 Item: Title: Results Destination: section.3 Children: Item: Title: Synthesis and Biophysical Properties of S-AuNPs-VLPs Destination: subsection.3.1 Item: Title: Evaluation of SARS-CoV-2-Specific Humoral and Cellular Immune Responses Induced by AuNP-VLPS Presenting the S Protein in BALB/c Mice Destination: subsection.3.2 Item: Title: Discussion Destination: section.4 Item: Title: Conclusions Destination: section.5 Item: Title: References Destination: appendix.A. Info: Title: Gold Nanoparticle Virus-like Particles Presenting SARS-CoV-2 Spike Protein: Synthesis, Biophysical Properties and Immunogenicity in BALB/c Mice Author: Vivian A. Salazar, Joan Comenge, Rosa Suárez-López, Judith A. Burger, Rogier W. Sanders, Neus G. Bastús, Carlos Jaime, Joan Joseph-Munne and Victor Puntes Subject: Gold nanoparticles (AuNPs) decorated with antigens have recently emerged as promising tools for vaccine development due to their innate ability to provide stability to antigens and modulate immune responses. In this study, we have engineered deactivated virus-like particles (VLPs) by precisely functionalizing gold cores with coronas comprising the full SARS-CoV-2 spike protein (S). Using BALB/c mice as a model, we investigated the immunogenicity of these S-AuNPs-VLPs. Our results demonstrate that S-AuNPs-VLPs consistently enhanced antigen-specific antibody responses compared to the S protein free in solution. This enhancement included higher binding antibody titers, higher neutralizing capacity of antibodies, and stronger T-cell responses. Compared to the mRNA COVID-19 vaccine, where the S protein is synthesized in situ, S-AuNPs-VLPs induced comparable binding and neutralizing antibody responses, but substantially superior T-cell responses. In conclusion, our study highlights the potential of conjugated AuNPs as an effective antigen-delivery system for protein-based vaccines targeting a broad spectrum of infectious diseases and other emergent viruses. 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