Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development
Background/Objectives: Foot-and-mouth disease virus (FMDV) poses a continuous threat to livestock health and agricultural economies. Current vaccines require high biosafety standards and are costly to produce. While novel vaccine technologies have been explored, most fail to meet industrial scalabil...
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| Format: | info:ar-repo/semantics/artículo |
| Language: | Inglés |
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MDPI
2025
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| Online Access: | http://hdl.handle.net/20.500.12123/22626 https://www.mdpi.com/2076-393X/13/6/581 https://doi.org/10.3390/vaccines13060581 |
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| author | Mignaqui, Ana Clara Ferella, Alejandra Sánchez, Cintia Stuible, Matthew Scian, Romina Filippi, Jorge Cardillo, Sabrina Beatriz Durocher, Yves Wigdorovitz, Andres |
| author_browse | Cardillo, Sabrina Beatriz Durocher, Yves Ferella, Alejandra Filippi, Jorge Mignaqui, Ana Clara Scian, Romina Stuible, Matthew Sánchez, Cintia Wigdorovitz, Andres |
| author_facet | Mignaqui, Ana Clara Ferella, Alejandra Sánchez, Cintia Stuible, Matthew Scian, Romina Filippi, Jorge Cardillo, Sabrina Beatriz Durocher, Yves Wigdorovitz, Andres |
| author_sort | Mignaqui, Ana Clara |
| collection | INTA Digital |
| description | Background/Objectives: Foot-and-mouth disease virus (FMDV) poses a continuous threat to livestock health and agricultural economies. Current vaccines require high biosafety standards and are costly to produce. While novel vaccine technologies have been explored, most fail to meet industrial scalability, cost-efficiency, or multiserotype flexibility required for effective FMD control. This study aimed to evaluate the feasibility of using a high-cell density transient gene expression (TGE) system in CHO cells for the production of FMDV virus-like particles (VLPs) as a recombinant vaccine platform. Methods: VLP expression was optimized by adjusting cDNA and polyethyleneimine (PEI) concentrations. Expression yields were compared at 24 and 48 h post-transfection to determine optimal harvest timing. We further tested the system’s capacity to express different serotypes and chimeric constructs, incorporating VP1 sequences from various FMDV strains. Immunogenicity was evaluated in swine using VLPs from the A2001 Argentina strain as a model. Results: Optimal VLP expression was achieved at 24 h post-transfection. Chimeric constructs incorporating heterologous VP1 regions were successfully expressed. Immunized pigs developed protective antibody titers as measured by a virus neutralization test (VNT, log10 titer 1.43) and liquid-phase blocking ELISA (LPBE, titer 2.20) at 28 days post-vaccination (dpv). Titers remained above protective thresholds up to 60 dpv with a single dose. A booster at 28 dpv further elevated titers to levels comparable to those induced by the inactivated vaccine. Conclusions: Our results demonstrate the feasibility of using CHO cell-based TGE for producing immunogenic FMDV VLPs. This platform shows promise for scalable, cost-effective, and biosafe development of recombinant FMD vaccines. |
| format | info:ar-repo/semantics/artículo |
| id | INTA22626 |
| institution | Instituto Nacional de Tecnología Agropecuaria (INTA -Argentina) |
| language | Inglés |
| publishDate | 2025 |
| publishDateRange | 2025 |
| publishDateSort | 2025 |
| publisher | MDPI |
| publisherStr | MDPI |
| record_format | dspace |
| spelling | INTA226262025-06-11T11:26:28Z Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development Mignaqui, Ana Clara Ferella, Alejandra Sánchez, Cintia Stuible, Matthew Scian, Romina Filippi, Jorge Cardillo, Sabrina Beatriz Durocher, Yves Wigdorovitz, Andres Virus-like Particles Foot-and-mouth Disease Aphthovirus Gene Expression Vaccines Partícula Similar a Virus Fiebre Aftosa Virus Fiebre Aftosa Expresión Génica Vacuna Background/Objectives: Foot-and-mouth disease virus (FMDV) poses a continuous threat to livestock health and agricultural economies. Current vaccines require high biosafety standards and are costly to produce. While novel vaccine technologies have been explored, most fail to meet industrial scalability, cost-efficiency, or multiserotype flexibility required for effective FMD control. This study aimed to evaluate the feasibility of using a high-cell density transient gene expression (TGE) system in CHO cells for the production of FMDV virus-like particles (VLPs) as a recombinant vaccine platform. Methods: VLP expression was optimized by adjusting cDNA and polyethyleneimine (PEI) concentrations. Expression yields were compared at 24 and 48 h post-transfection to determine optimal harvest timing. We further tested the system’s capacity to express different serotypes and chimeric constructs, incorporating VP1 sequences from various FMDV strains. Immunogenicity was evaluated in swine using VLPs from the A2001 Argentina strain as a model. Results: Optimal VLP expression was achieved at 24 h post-transfection. Chimeric constructs incorporating heterologous VP1 regions were successfully expressed. Immunized pigs developed protective antibody titers as measured by a virus neutralization test (VNT, log10 titer 1.43) and liquid-phase blocking ELISA (LPBE, titer 2.20) at 28 days post-vaccination (dpv). Titers remained above protective thresholds up to 60 dpv with a single dose. A booster at 28 dpv further elevated titers to levels comparable to those induced by the inactivated vaccine. Conclusions: Our results demonstrate the feasibility of using CHO cell-based TGE for producing immunogenic FMDV VLPs. This platform shows promise for scalable, cost-effective, and biosafe development of recombinant FMD vaccines. Instituto de Virología Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas. INCUINTA; Argentina Fil: Mignaqui, Ana Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina Fil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; Argentina Fil: Ferella, Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas. INCUINTA; Argentina Fil: Ferella, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina Fil: Sánchez, Cintia. Biogénesis Bagó; Argentina Fil: Stuible, Matthew. National Research Council Canada. Human Health Therapeutics Research Center; Canadá Fil: Scian, Romina. Biogénesis Bagó; Argentina Fil: Filippi, Jorge. Biogénesis Bagó; Argentina Fil: Cardillo, Sabrina Beatriz. Biogénesis Bagó; Argentina Fil: Durocher, Yves. National Research Council Canada. Human Health Therapeutics Research Center; Canadá Fil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas. INCUINTA; Argentina Fil: Wigdorovitz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina 2025-06-11T10:45:04Z 2025-06-11T10:45:04Z 2025-06 info:ar-repo/semantics/artículo info:eu-repo/semantics/article info:eu-repo/semantics/publishedVersion http://hdl.handle.net/20.500.12123/22626 https://www.mdpi.com/2076-393X/13/6/581 2076-393X https://doi.org/10.3390/vaccines13060581 eng info:eu-repograntAgreement/INTA/2019-PD-E5-I105-001, Patógenos animales: su interacción con el hospedador y el medio ambiente. Impacto en productividad, ecosistemas, sanidad animal y salud pública en el marco ?Una Salud? info:eu-repo/semantics/openAccess http://creativecommons.org/licenses/by-nc-sa/4.0/ Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) application/pdf MDPI Vaccines 13 (6) : 581 (June 2025) |
| spellingShingle | Virus-like Particles Foot-and-mouth Disease Aphthovirus Gene Expression Vaccines Partícula Similar a Virus Fiebre Aftosa Virus Fiebre Aftosa Expresión Génica Vacuna Mignaqui, Ana Clara Ferella, Alejandra Sánchez, Cintia Stuible, Matthew Scian, Romina Filippi, Jorge Cardillo, Sabrina Beatriz Durocher, Yves Wigdorovitz, Andres Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development |
| title | Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development |
| title_full | Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development |
| title_fullStr | Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development |
| title_full_unstemmed | Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development |
| title_short | Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development |
| title_sort | optimized production of virus like particles in a high cho cell density transient gene expression system for foot and mouth disease vaccine development |
| topic | Virus-like Particles Foot-and-mouth Disease Aphthovirus Gene Expression Vaccines Partícula Similar a Virus Fiebre Aftosa Virus Fiebre Aftosa Expresión Génica Vacuna |
| url | http://hdl.handle.net/20.500.12123/22626 https://www.mdpi.com/2076-393X/13/6/581 https://doi.org/10.3390/vaccines13060581 |
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