Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo

Autores
Gómez Sebastián, Silvia; Nuñez, Maria C.; Garaicoechea, Lorena Laura; Alvarado, Carmen; Mozgovoj, Marina Valeria; Lasa, Rodrigo; Kahl, Alan Jonatan; Wigdorovitz, Andrés; Parreño, Gladys Viviana; Escribano, José M.
Año de publicación
2012
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Background: Single-domain antibodies (sdAbs), also known as nanobodies or VHHs, are characterized by high stability and solubility, thus maintaining the affinity and therapeutic value provided by conventional antibodies. Given these properties, VHHs offer a novel alternative to classical antibody approaches. To date, VHHs have been produced mainly in E. coli, yeast, plants and mammalian cells. To apply the single-domain antibodies as a preventive or therapeutic strategy to control rotavirus infections in developing countries (444,000 deaths in children under 5 years of age) has to be minimized their production costs. Results: Here we describe the highly efficient expression of functional VHHs by the Improved Baculovirus Expression System (IBESW technology), which uses a baculovirus expression vector in combination with Trichoplusia ni larvae as living biofactories. Two VHHs, named 3B2 and 2KD1, specific for the inner capsid protein VP6 of Group A rotavirus, were expressed in insect larvae. The IBESW technology achieved very high expression of 3B2 and 2KD1, reaching 2.62% and 3.63% of the total soluble protein obtained from larvae, respectively. These expression levels represent up to 257 mg/L of protein extract after insect processing (1 L extract represents about 125 g of insect biomass or about 375 insect larvae). Larva-derived antibodies were fully functional when tested in vitro and in vivo, neutralizing Group A rotaviruses and protecting offspring mice against rotavirus-induced diarrhea. Conclusions: Our results open up the possibility of using insects as living biofactories (IBESW technology) for the cost-efficient production of these and other fully functional VHHs to be used for diagnostic or therapeutic purposes, thereby eliminating concerns regarding the use of bacterial or mammalian cells. To the best of our knowledge, this is the first time that insects have been used as living biofactories to produce a VHH molecule.
Fil: Gómez Sebastián, Silvia. Universidad Politécnica de Madrid; España
Fil: Nuñez, Maria C.. Universidad Politécnica de Madrid; España
Fil: Garaicoechea, Lorena Laura. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alvarado, Carmen. Universidad Politécnica de Madrid; España
Fil: Mozgovoj, Marina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Lasa, Rodrigo. Universidad Politécnica de Madrid; España
Fil: Kahl, Alan Jonatan. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Wigdorovitz, Andrés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Escribano, José M.. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
Materia
SINGLE-DOMAIN ANTIBODIES
THERAPEUTIC MOLECULE
NEUTRALIZATION
ROTAVIRUS A
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/269299
Acceder
id CONICETDig_0b2bbfa4641774e8aa81cbbf5cd0c4a3
oai_identifier_str oai:ri.conicet.gov.ar:11336/269299
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivoGómez Sebastián, SilviaNuñez, Maria C.Garaicoechea, Lorena LauraAlvarado, CarmenMozgovoj, Marina ValeriaLasa, RodrigoKahl, Alan JonatanWigdorovitz, AndrésParreño, Gladys VivianaEscribano, José M.SINGLE-DOMAIN ANTIBODIESTHERAPEUTIC MOLECULENEUTRALIZATIONROTAVIRUS Ahttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Background: Single-domain antibodies (sdAbs), also known as nanobodies or VHHs, are characterized by high stability and solubility, thus maintaining the affinity and therapeutic value provided by conventional antibodies. Given these properties, VHHs offer a novel alternative to classical antibody approaches. To date, VHHs have been produced mainly in E. coli, yeast, plants and mammalian cells. To apply the single-domain antibodies as a preventive or therapeutic strategy to control rotavirus infections in developing countries (444,000 deaths in children under 5 years of age) has to be minimized their production costs. Results: Here we describe the highly efficient expression of functional VHHs by the Improved Baculovirus Expression System (IBESW technology), which uses a baculovirus expression vector in combination with Trichoplusia ni larvae as living biofactories. Two VHHs, named 3B2 and 2KD1, specific for the inner capsid protein VP6 of Group A rotavirus, were expressed in insect larvae. The IBESW technology achieved very high expression of 3B2 and 2KD1, reaching 2.62% and 3.63% of the total soluble protein obtained from larvae, respectively. These expression levels represent up to 257 mg/L of protein extract after insect processing (1 L extract represents about 125 g of insect biomass or about 375 insect larvae). Larva-derived antibodies were fully functional when tested in vitro and in vivo, neutralizing Group A rotaviruses and protecting offspring mice against rotavirus-induced diarrhea. Conclusions: Our results open up the possibility of using insects as living biofactories (IBESW technology) for the cost-efficient production of these and other fully functional VHHs to be used for diagnostic or therapeutic purposes, thereby eliminating concerns regarding the use of bacterial or mammalian cells. To the best of our knowledge, this is the first time that insects have been used as living biofactories to produce a VHH molecule.Fil: Gómez Sebastián, Silvia. Universidad Politécnica de Madrid; EspañaFil: Nuñez, Maria C.. Universidad Politécnica de Madrid; EspañaFil: Garaicoechea, Lorena Laura. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Alvarado, Carmen. Universidad Politécnica de Madrid; EspañaFil: Mozgovoj, Marina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; ArgentinaFil: Lasa, Rodrigo. Universidad Politécnica de Madrid; EspañaFil: Kahl, Alan Jonatan. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Wigdorovitz, Andrés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; ArgentinaFil: Escribano, José M.. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; EspañaBioMed Central2012-09info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/269299Gómez Sebastián, Silvia; Nuñez, Maria C.; Garaicoechea, Lorena Laura; Alvarado, Carmen; Mozgovoj, Marina Valeria; et al.; Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo; BioMed Central; Bmc Biotechnology; 12; 1; 9-2012; 59-691472-6750CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.biomedcentral.com/1472-6750/12/59info:eu-repo/semantics/altIdentifier/doi/10.1186/1472-6750-12-59info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:48:18Zoai:ri.conicet.gov.ar:11336/269299instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-09-03 09:48:18.792CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
title Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
spellingShingle Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
Gómez Sebastián, Silvia
SINGLE-DOMAIN ANTIBODIES
THERAPEUTIC MOLECULE
NEUTRALIZATION
ROTAVIRUS A
title_short Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
title_full Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
title_fullStr Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
title_full_unstemmed Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
title_sort Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo
dc.creator.none.fl_str_mv Gómez Sebastián, Silvia
Nuñez, Maria C.
Garaicoechea, Lorena Laura
Alvarado, Carmen
Mozgovoj, Marina Valeria
Lasa, Rodrigo
Kahl, Alan Jonatan
Wigdorovitz, Andrés
Parreño, Gladys Viviana
Escribano, José M.
author Gómez Sebastián, Silvia
author_facet Gómez Sebastián, Silvia
Nuñez, Maria C.
Garaicoechea, Lorena Laura
Alvarado, Carmen
Mozgovoj, Marina Valeria
Lasa, Rodrigo
Kahl, Alan Jonatan
Wigdorovitz, Andrés
Parreño, Gladys Viviana
Escribano, José M.
author_role author
author2 Nuñez, Maria C.
Garaicoechea, Lorena Laura
Alvarado, Carmen
Mozgovoj, Marina Valeria
Lasa, Rodrigo
Kahl, Alan Jonatan
Wigdorovitz, Andrés
Parreño, Gladys Viviana
Escribano, José M.
author2_role author
author
author
author
author
author
author
author
author
dc.subject.none.fl_str_mv SINGLE-DOMAIN ANTIBODIES
THERAPEUTIC MOLECULE
NEUTRALIZATION
ROTAVIRUS A
topic SINGLE-DOMAIN ANTIBODIES
THERAPEUTIC MOLECULE
NEUTRALIZATION
ROTAVIRUS A
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Background: Single-domain antibodies (sdAbs), also known as nanobodies or VHHs, are characterized by high stability and solubility, thus maintaining the affinity and therapeutic value provided by conventional antibodies. Given these properties, VHHs offer a novel alternative to classical antibody approaches. To date, VHHs have been produced mainly in E. coli, yeast, plants and mammalian cells. To apply the single-domain antibodies as a preventive or therapeutic strategy to control rotavirus infections in developing countries (444,000 deaths in children under 5 years of age) has to be minimized their production costs. Results: Here we describe the highly efficient expression of functional VHHs by the Improved Baculovirus Expression System (IBESW technology), which uses a baculovirus expression vector in combination with Trichoplusia ni larvae as living biofactories. Two VHHs, named 3B2 and 2KD1, specific for the inner capsid protein VP6 of Group A rotavirus, were expressed in insect larvae. The IBESW technology achieved very high expression of 3B2 and 2KD1, reaching 2.62% and 3.63% of the total soluble protein obtained from larvae, respectively. These expression levels represent up to 257 mg/L of protein extract after insect processing (1 L extract represents about 125 g of insect biomass or about 375 insect larvae). Larva-derived antibodies were fully functional when tested in vitro and in vivo, neutralizing Group A rotaviruses and protecting offspring mice against rotavirus-induced diarrhea. Conclusions: Our results open up the possibility of using insects as living biofactories (IBESW technology) for the cost-efficient production of these and other fully functional VHHs to be used for diagnostic or therapeutic purposes, thereby eliminating concerns regarding the use of bacterial or mammalian cells. To the best of our knowledge, this is the first time that insects have been used as living biofactories to produce a VHH molecule.
Fil: Gómez Sebastián, Silvia. Universidad Politécnica de Madrid; España
Fil: Nuñez, Maria C.. Universidad Politécnica de Madrid; España
Fil: Garaicoechea, Lorena Laura. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Alvarado, Carmen. Universidad Politécnica de Madrid; España
Fil: Mozgovoj, Marina Valeria. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina
Fil: Lasa, Rodrigo. Universidad Politécnica de Madrid; España
Fil: Kahl, Alan Jonatan. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Wigdorovitz, Andrés. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Parreño, Gladys Viviana. Instituto Nacional de Tecnología Agropecuaria. Centro de Investigación en Ciencias Veterinarias y Agronómicas. Instituto de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina
Fil: Escribano, José M.. Instituto Nacional de Investigación y Tecnología Agraria y Alimentaria; España
description Background: Single-domain antibodies (sdAbs), also known as nanobodies or VHHs, are characterized by high stability and solubility, thus maintaining the affinity and therapeutic value provided by conventional antibodies. Given these properties, VHHs offer a novel alternative to classical antibody approaches. To date, VHHs have been produced mainly in E. coli, yeast, plants and mammalian cells. To apply the single-domain antibodies as a preventive or therapeutic strategy to control rotavirus infections in developing countries (444,000 deaths in children under 5 years of age) has to be minimized their production costs. Results: Here we describe the highly efficient expression of functional VHHs by the Improved Baculovirus Expression System (IBESW technology), which uses a baculovirus expression vector in combination with Trichoplusia ni larvae as living biofactories. Two VHHs, named 3B2 and 2KD1, specific for the inner capsid protein VP6 of Group A rotavirus, were expressed in insect larvae. The IBESW technology achieved very high expression of 3B2 and 2KD1, reaching 2.62% and 3.63% of the total soluble protein obtained from larvae, respectively. These expression levels represent up to 257 mg/L of protein extract after insect processing (1 L extract represents about 125 g of insect biomass or about 375 insect larvae). Larva-derived antibodies were fully functional when tested in vitro and in vivo, neutralizing Group A rotaviruses and protecting offspring mice against rotavirus-induced diarrhea. Conclusions: Our results open up the possibility of using insects as living biofactories (IBESW technology) for the cost-efficient production of these and other fully functional VHHs to be used for diagnostic or therapeutic purposes, thereby eliminating concerns regarding the use of bacterial or mammalian cells. To the best of our knowledge, this is the first time that insects have been used as living biofactories to produce a VHH molecule.
publishDate 2012
dc.date.none.fl_str_mv 2012-09
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/11336/269299
Gómez Sebastián, Silvia; Nuñez, Maria C.; Garaicoechea, Lorena Laura; Alvarado, Carmen; Mozgovoj, Marina Valeria; et al.; Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo; BioMed Central; Bmc Biotechnology; 12; 1; 9-2012; 59-69
1472-6750
CONICET Digital
CONICET
url http://hdl.handle.net/11336/269299
identifier_str_mv Gómez Sebastián, Silvia; Nuñez, Maria C.; Garaicoechea, Lorena Laura; Alvarado, Carmen; Mozgovoj, Marina Valeria; et al.; Rotavirus A-specific single-domain antibodies produced in baculovirus-infected insect larvae are protective in vivo; BioMed Central; Bmc Biotechnology; 12; 1; 9-2012; 59-69
1472-6750
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/http://www.biomedcentral.com/1472-6750/12/59
info:eu-repo/semantics/altIdentifier/doi/10.1186/1472-6750-12-59
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv BioMed Central
publisher.none.fl_str_mv BioMed Central
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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