bc54042cd03fa969531c5eb10ce8cc94 vaccines-11-01641.pdf 4c52b7462b8909036d3694fab201ae04e618947b vaccines-11-01641.pdf 16bd6a845973b4dd71359d20d363e40c33e22f374df111d7034af0e9e0374318 vaccines-11-01641.pdf Title: Gag Virus-like Particles Functionalized with SARS-CoV-2 Variants: Generation, Characterization and Recognition by COVID-19 Convalescent Patients’ Sera Subject: The robustness, safety, versatility, and high immunogenicity of virus-like particles (VLPs) make them a promising approach for the generation of vaccines against a broad range of pathogens. VLPs are recombinant macromolecular structures that closely mimic the native conformation of viruses without carrying viral genetic material. Particularly, HIV-1 Gag-based VLPs are a suitable platform for the presentation of the SARS-CoV-2 Spike (S) protein on their surface. In this context, this work studies the effect of different rationally engineered mutations of the S protein to improve some of its characteristics. The studied variants harbored mutations such as proline substitutions for S stabilization, D614G from the early dominant pandemic form, the elimination of the S1/S2 furin cleavage site to improve S homogeneity, the suppression of a retention motif to favor its membrane localization, and cysteine substitutions to increase its immunogenicity and avoid potential undesired antibody-dependent enhancement (ADE) effects. The influence of the mutations on VLP expression was studied, as well as their immunogenic potential, by testing the recognition of the generated VLP variants by COVID-19 convalescent patients’ sera. The results of this work are conceived to give insights on the selection of S protein candidates for their use as immunogens and to showcase the potential of VLPs as carriers for antigen presentation. Keywords: SARS-CoV-2; variants; spike; virus-like particles; COVID-19; D614G; furin cleavage Author: Arnau Boix-Besora, Francesc Gòdia and Laura Cervera Creator: LaTeX with hyperref Producer: pdfTeX-1.40.21 CreationDate: Thu Oct 26 08:02:20 2023 CEST ModDate: Thu Oct 26 08:05:48 2023 CEST Tagged: no UserProperties: no Suspects: no Form: none JavaScript: no Pages: 16 Encrypted: no Page size: 595.276 x 841.89 pts (A4) Page rot: 0 File size: 2493141 bytes Optimized: no PDF version: 1.7 name type encoding emb sub uni object ID ------------------------------------ ----------------- ---------------- --- --- --- --------- WLFTRW+VnURWPalladioL-Bold Type 1 Custom yes yes no 10 0 GKYVXB+VnURWPalladioL Type 1 Custom yes yes no 15 0 WJNTVD+URWPalladioL-Bold Type 1 Custom yes yes no 19 0 OFXJJX+URWPalladioL-Roma Type 1 Custom yes yes no 24 0 FVJXLY+URWPalladioL-Ital Type 1 Custom yes yes no 28 0 OJYQXC+CMSY10 Type 1 Builtin yes yes no 65 0 KICFIE+PalatinoLinotype,Bold TrueType WinAnsi yes yes no 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edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4a87761d Item: Title: Furin Cleavage Site Removal Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@66d1af89 Item: Title: Dibasic Motif Removal Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@8646db9 Item: Title: S-VLPs Variant Production and Quantification Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@37374a5e Item: Title: Assessment of the Immunogenic Potential of the S-VLP Candidates Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@4671e53b Item: Title: Discussion Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@2db7a79b Item: Title: References Destination: edu.harvard.hul.ois.jhove.module.pdf.PdfSimpleObject@6950e31 Info: Title: Gag Virus-like Particles Functionalized with SARS-CoV-2 Variants: Generation, Characterization and Recognition by COVID-19 Convalescent Patients’ Sera Author: Arnau Boix-Besora, Francesc Gòdia and Laura Cervera Subject: The robustness, safety, versatility, and high immunogenicity of virus-like particles (VLPs) make them a promising approach for the generation of vaccines against a broad range of pathogens. VLPs are recombinant macromolecular structures that closely mimic the native conformation of viruses without carrying viral genetic material. Particularly, HIV-1 Gag-based VLPs are a suitable platform for the presentation of the SARS-CoV-2 Spike (S) protein on their surface. In this context, this work studies the effect of different rationally engineered mutations of the S protein to improve some of its characteristics. The studied variants harbored mutations such as proline substitutions for S stabilization, D614G from the early dominant pandemic form, the elimination of the S1/S2 furin cleavage site to improve S homogeneity, the suppression of a retention motif to favor its membrane localization, and cysteine substitutions to increase its immunogenicity and avoid potential undesired antibody-dependent enhancement (ADE) effects. The influence of the mutations on VLP expression was studied, as well as their immunogenic potential, by testing the recognition of the generated VLP variants by COVID-19 convalescent patients’ sera. The results of this work are conceived to give insights on the selection of S protein candidates for their use as immunogens and to showcase the potential of VLPs as carriers for antigen presentation. 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