Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development

Autores
Mignaqui, Ana Clara; Ferella, Alejandra; Sánchez, Cintia; Stuible, Matthew; Scian, Romina; Filippi, Jorge; Cardillo, Sabrina Beatriz; Durocher, Yves; Wigdorovitz, Andres
Año de publicación
2025
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
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
Fuente
Vaccines 13 (6) : 581 (June 2025)
Materia
Virus-like Particles
Foot-and-mouth Disease
Aphthovirus
Gene Expression
Vaccines
Partícula Similar a Virus
Fiebre Aftosa
Virus Fiebre Aftosa
Expresión Génica
Vacuna
Nivel de accesibilidad
acceso abierto
Condiciones de uso
http://creativecommons.org/licenses/by-nc-sa/4.0/
Repositorio
INTA Digital (INTA)
Institución
Instituto Nacional de Tecnología Agropecuaria
OAI Identificador
oai:localhost:20.500.12123/22626
Acceder
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oai_identifier_str oai:localhost:20.500.12123/22626
network_acronym_str INTADig
repository_id_str l
network_name_str INTA Digital (INTA)
spelling Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine developmentMignaqui, Ana ClaraFerella, AlejandraSánchez, CintiaStuible, MatthewScian, RominaFilippi, JorgeCardillo, Sabrina BeatrizDurocher, YvesWigdorovitz, AndresVirus-like ParticlesFoot-and-mouth DiseaseAphthovirusGene ExpressionVaccinesPartícula Similar a VirusFiebre AftosaVirus Fiebre AftosaExpresión GénicaVacunaBackground/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 VirologíaFil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas. INCUINTA; ArgentinaFil: Mignaqui, Ana Clara. Consejo Nacional de Investigaciones Científicas y Técnicas. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; ArgentinaFil: Mignaqui, Ana Clara. Instituto Nacional de Tecnología Agropecuaria (INTA). Estación Experimental Agropecuaria Bariloche. Instituto de Investigaciones Forestales y Agropecuarias Bariloche; ArgentinaFil: Ferella, Alejandra. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas. INCUINTA; ArgentinaFil: Ferella, Alejandra. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Sánchez, Cintia. Biogénesis Bagó; ArgentinaFil: Stuible, Matthew. National Research Council Canada. Human Health Therapeutics Research Center; CanadáFil: Scian, Romina. Biogénesis Bagó; ArgentinaFil: Filippi, Jorge. Biogénesis Bagó; ArgentinaFil: Cardillo, Sabrina Beatriz. Biogénesis Bagó; ArgentinaFil: Durocher, Yves. National Research Council Canada. Human Health Therapeutics Research Center; CanadáFil: Wigdorovitz, Andres. Instituto Nacional de Tecnología Agropecuaria (INTA). Instituto de Virología e Innovaciones Tecnológicas. INCUINTA; ArgentinaFil: Wigdorovitz, Andres. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaMDPI2025-06-11T10:45:04Z2025-06-11T10:45:04Z2025-06info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfhttp://hdl.handle.net/20.500.12123/22626https://www.mdpi.com/2076-393X/13/6/5812076-393Xhttps://doi.org/10.3390/vaccines13060581Vaccines 13 (6) : 581 (June 2025)reponame:INTA Digital (INTA)instname:Instituto Nacional de Tecnología Agropecuariaenginfo: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/openAccesshttp://creativecommons.org/licenses/by-nc-sa/4.0/Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)2025-09-29T13:47:21Zoai:localhost:20.500.12123/22626instacron:INTAInstitucionalhttp://repositorio.inta.gob.ar/Organismo científico-tecnológicoNo correspondehttp://repositorio.inta.gob.ar/oai/requesttripaldi.nicolas@inta.gob.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:l2025-09-29 13:47:21.464INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuariafalse
dc.title.none.fl_str_mv 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
spellingShingle 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
Virus-like Particles
Foot-and-mouth Disease
Aphthovirus
Gene Expression
Vaccines
Partícula Similar a Virus
Fiebre Aftosa
Virus Fiebre Aftosa
Expresión Génica
Vacuna
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_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_sort Optimized production of virus-like particles in a high-CHO-cell-density transient gene expression system for foot-and-mouth disease vaccine development
dc.creator.none.fl_str_mv Mignaqui, Ana Clara
Ferella, Alejandra
Sánchez, Cintia
Stuible, Matthew
Scian, Romina
Filippi, Jorge
Cardillo, Sabrina Beatriz
Durocher, Yves
Wigdorovitz, Andres
author Mignaqui, Ana Clara
author_facet Mignaqui, Ana Clara
Ferella, Alejandra
Sánchez, Cintia
Stuible, Matthew
Scian, Romina
Filippi, Jorge
Cardillo, Sabrina Beatriz
Durocher, Yves
Wigdorovitz, Andres
author_role author
author2 Ferella, Alejandra
Sánchez, Cintia
Stuible, Matthew
Scian, Romina
Filippi, Jorge
Cardillo, Sabrina Beatriz
Durocher, Yves
Wigdorovitz, Andres
author2_role author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv Virus-like Particles
Foot-and-mouth Disease
Aphthovirus
Gene Expression
Vaccines
Partícula Similar a Virus
Fiebre Aftosa
Virus Fiebre Aftosa
Expresión Génica
Vacuna
topic Virus-like Particles
Foot-and-mouth Disease
Aphthovirus
Gene Expression
Vaccines
Partícula Similar a Virus
Fiebre Aftosa
Virus Fiebre Aftosa
Expresión Génica
Vacuna
dc.description.none.fl_txt_mv 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
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.
publishDate 2025
dc.date.none.fl_str_mv 2025-06-11T10:45:04Z
2025-06-11T10:45:04Z
2025-06
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv 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
url http://hdl.handle.net/20.500.12123/22626
https://www.mdpi.com/2076-393X/13/6/581
https://doi.org/10.3390/vaccines13060581
identifier_str_mv 2076-393X
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv 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?
dc.rights.none.fl_str_mv 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)
eu_rights_str_mv openAccess
rights_invalid_str_mv http://creativecommons.org/licenses/by-nc-sa/4.0/
Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0)
dc.format.none.fl_str_mv application/pdf
dc.publisher.none.fl_str_mv MDPI
publisher.none.fl_str_mv MDPI
dc.source.none.fl_str_mv Vaccines 13 (6) : 581 (June 2025)
reponame:INTA Digital (INTA)
instname:Instituto Nacional de Tecnología Agropecuaria
reponame_str INTA Digital (INTA)
collection INTA Digital (INTA)
instname_str Instituto Nacional de Tecnología Agropecuaria
repository.name.fl_str_mv INTA Digital (INTA) - Instituto Nacional de Tecnología Agropecuaria
repository.mail.fl_str_mv tripaldi.nicolas@inta.gob.ar
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