Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source

Autores
Arias, Cesar Andres Diaz; Marques, Daniela de Araujo Viana; Pellegrini Malpiedi, Luciana; Maranhão, Andrea Queiroz; Abdalla, Dulcinéia Saes Parra; Converti, Attilio; Junior, Adalberto Pessoa
Año de publicación
2017
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (−) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric productivity of the antibody fragment secreted by cells precultured, cryopreserved and reactivated in glycerol was 42.9% higher compared with cells precultured in glucose, the use of glycerol in bioreactor led to a remarkable shortening of the lag phase, thereby increasing it by no less than thrice compared to flasks. These results are quite promising in comparison with those reported in the literature for possible future industrial applications of this cultivation, taking into account that the overall process time was reduced by around 8 h.
Fil: Arias, Cesar Andres Diaz. Universidade de Sao Paulo; Brasil
Fil: Marques, Daniela de Araujo Viana. Universidade Federal Rural Pernambuco; Brasil
Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidade de Sao Paulo; Brasil
Fil: Maranhão, Andrea Queiroz. Universidade Nacional de Brasíilia; Brasil
Fil: Abdalla, Dulcinéia Saes Parra. Universidade de Sao Paulo; Brasil
Fil: Converti, Attilio. University of Genoa; Italia
Fil: Junior, Adalberto Pessoa. Universidade de Sao Paulo; Brasil
Materia
Carbon Source
Cryopreservation
Expression
Pichia Pastoris
Scfv Antibody Fragment
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/53280
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/53280
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network_name_str CONICET Digital (CONICET)
spelling Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon sourceArias, Cesar Andres DiazMarques, Daniela de Araujo VianaPellegrini Malpiedi, LucianaMaranhão, Andrea QueirozAbdalla, Dulcinéia Saes ParraConverti, AttilioJunior, Adalberto PessoaCarbon SourceCryopreservationExpressionPichia PastorisScfv Antibody Fragmenthttps://purl.org/becyt/ford/2.9https://purl.org/becyt/ford/2Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (−) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric productivity of the antibody fragment secreted by cells precultured, cryopreserved and reactivated in glycerol was 42.9% higher compared with cells precultured in glucose, the use of glycerol in bioreactor led to a remarkable shortening of the lag phase, thereby increasing it by no less than thrice compared to flasks. These results are quite promising in comparison with those reported in the literature for possible future industrial applications of this cultivation, taking into account that the overall process time was reduced by around 8 h.Fil: Arias, Cesar Andres Diaz. Universidade de Sao Paulo; BrasilFil: Marques, Daniela de Araujo Viana. Universidade Federal Rural Pernambuco; BrasilFil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidade de Sao Paulo; BrasilFil: Maranhão, Andrea Queiroz. Universidade Nacional de Brasíilia; BrasilFil: Abdalla, Dulcinéia Saes Parra. Universidade de Sao Paulo; BrasilFil: Converti, Attilio. University of Genoa; ItaliaFil: Junior, Adalberto Pessoa. Universidade de Sao Paulo; BrasilSociedade Brasileira de Microbiologia2017-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/53280Arias, Cesar Andres Diaz; Marques, Daniela de Araujo Viana; Pellegrini Malpiedi, Luciana; Maranhão, Andrea Queiroz; Abdalla, Dulcinéia Saes Parra; et al.; Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source; Sociedade Brasileira de Microbiologia; Brazilian Journal of Microbiology; 48; 3; 7-2017; 419-4261678-4405CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.bjm.2016.11.009info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S1517838216301769info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:03:24Zoai:ri.conicet.gov.ar:11336/53280instacron: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:34982025-09-29 10:03:24.917CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
title Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
spellingShingle Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
Arias, Cesar Andres Diaz
Carbon Source
Cryopreservation
Expression
Pichia Pastoris
Scfv Antibody Fragment
title_short Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
title_full Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
title_fullStr Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
title_full_unstemmed Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
title_sort Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source
dc.creator.none.fl_str_mv Arias, Cesar Andres Diaz
Marques, Daniela de Araujo Viana
Pellegrini Malpiedi, Luciana
Maranhão, Andrea Queiroz
Abdalla, Dulcinéia Saes Parra
Converti, Attilio
Junior, Adalberto Pessoa
author Arias, Cesar Andres Diaz
author_facet Arias, Cesar Andres Diaz
Marques, Daniela de Araujo Viana
Pellegrini Malpiedi, Luciana
Maranhão, Andrea Queiroz
Abdalla, Dulcinéia Saes Parra
Converti, Attilio
Junior, Adalberto Pessoa
author_role author
author2 Marques, Daniela de Araujo Viana
Pellegrini Malpiedi, Luciana
Maranhão, Andrea Queiroz
Abdalla, Dulcinéia Saes Parra
Converti, Attilio
Junior, Adalberto Pessoa
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv Carbon Source
Cryopreservation
Expression
Pichia Pastoris
Scfv Antibody Fragment
topic Carbon Source
Cryopreservation
Expression
Pichia Pastoris
Scfv Antibody Fragment
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.9
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (−) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric productivity of the antibody fragment secreted by cells precultured, cryopreserved and reactivated in glycerol was 42.9% higher compared with cells precultured in glucose, the use of glycerol in bioreactor led to a remarkable shortening of the lag phase, thereby increasing it by no less than thrice compared to flasks. These results are quite promising in comparison with those reported in the literature for possible future industrial applications of this cultivation, taking into account that the overall process time was reduced by around 8 h.
Fil: Arias, Cesar Andres Diaz. Universidade de Sao Paulo; Brasil
Fil: Marques, Daniela de Araujo Viana. Universidade Federal Rural Pernambuco; Brasil
Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Científicas y Técnicas; Argentina. Universidade de Sao Paulo; Brasil
Fil: Maranhão, Andrea Queiroz. Universidade Nacional de Brasíilia; Brasil
Fil: Abdalla, Dulcinéia Saes Parra. Universidade de Sao Paulo; Brasil
Fil: Converti, Attilio. University of Genoa; Italia
Fil: Junior, Adalberto Pessoa. Universidade de Sao Paulo; Brasil
description Antibodies and antibody fragments are nowadays among the most important biotechnological products, and Pichia pastoris is one of the most important vectors to produce them as well as other recombinant proteins. The conditions to effectively cultivate a P. pastoris strain previously genetically modified to produce the single-chain variable fragment anti low density lipoprotein (−) under the control of the alcohol oxidase promoter have been investigated in this study. In particular, it was evaluated if, and eventually how, the carbon source (glucose or glycerol) used in the preculture preceding cryopreservation in 20% glycerol influences both cell and antibody fragment productions either in flasks or in bioreactor. Although in flasks the volumetric productivity of the antibody fragment secreted by cells precultured, cryopreserved and reactivated in glycerol was 42.9% higher compared with cells precultured in glucose, the use of glycerol in bioreactor led to a remarkable shortening of the lag phase, thereby increasing it by no less than thrice compared to flasks. These results are quite promising in comparison with those reported in the literature for possible future industrial applications of this cultivation, taking into account that the overall process time was reduced by around 8 h.
publishDate 2017
dc.date.none.fl_str_mv 2017-07
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/11336/53280
Arias, Cesar Andres Diaz; Marques, Daniela de Araujo Viana; Pellegrini Malpiedi, Luciana; Maranhão, Andrea Queiroz; Abdalla, Dulcinéia Saes Parra; et al.; Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source; Sociedade Brasileira de Microbiologia; Brazilian Journal of Microbiology; 48; 3; 7-2017; 419-426
1678-4405
CONICET Digital
CONICET
url http://hdl.handle.net/11336/53280
identifier_str_mv Arias, Cesar Andres Diaz; Marques, Daniela de Araujo Viana; Pellegrini Malpiedi, Luciana; Maranhão, Andrea Queiroz; Abdalla, Dulcinéia Saes Parra; et al.; Cultivation of Pichia pastoris carrying the scFv anti LDL (−) antibody fragment. Effect of preculture carbon source; Sociedade Brasileira de Microbiologia; Brazilian Journal of Microbiology; 48; 3; 7-2017; 419-426
1678-4405
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.bjm.2016.11.009
info:eu-repo/semantics/altIdentifier/url/https://linkinghub.elsevier.com/retrieve/pii/S1517838216301769
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-nd/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
dc.publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
publisher.none.fl_str_mv Sociedade Brasileira de Microbiologia
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 13.070432

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