Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification

Autores
Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Abdalla, Dulcinéia Saes Parra; de Alcantara Pessoa, Pedro; Pessoa, Adalberto Jr
Año de publicación
2014
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The main goal of this work was to evaluate the feasibility of using aqueous micellar two-phase systems (AMTPS) of Triton X-114 to purify a single-chain antibody fragment (scFv) directly from yeast fermentation supernatant. The coexistence curves of aqueous micellar two-phase systems, highly loaded (up to 90% wt/wt) with the biological feedstock, were determined. Besides, the effect of several additives such as inorganic salts and affinity ligands on the phase separation behavior was investigated. The obtained coexistence curves demonstrated that the assayed surfactant/yeast broth solutions were able to separate into two phases at temperatures lower than 24 C. This information was then utilized to assay scFv partitioning and purification. In general, proteins present in the yeast broth, including the scFv, partitioned to the top, micelle-depleted phase due to its hydrophilic character. When affinity ligands were used, an opposite behavior was observed for scFv due to the uneven partitioning of ligands toward the micellarrich phase. The best purification performances were attained for the system consisting of 4% wt/wt of Triton X-114, 60% wt/wt of yeast fermentation supernatant and the synthetic ligand Fabsorbent™ F1P HF, with a recovery percentage of 88% and a purification factor of 2. These results demonstrate the potential applicability of these systems for designing early steps for scFv purification directly from yeast broth. New perspectives are now open for the use of this methodology for recombinant antibody fragments purification
Fil: Pellegrini Malpiedi, Luciana. Universidade de Sao Paulo; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Quimicos Rosario. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Instituto de Procesos Biotecnológicos y Quimicos Rosario; Argentina
Fil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Quimicos Rosario. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Instituto de Procesos Biotecnológicos y Quimicos Rosario; Argentina
Fil: Abdalla, Dulcinéia Saes Parra. Universidade de Sao Paulo; Brasil
Fil: de Alcantara Pessoa, Pedro. Universidade de Sao Paulo; Brasil
Fil: Pessoa, Adalberto Jr. Universidade de Sao Paulo; Brasil
Materia
Aqueous Micellar Two-Phase System
Cloud Point
Clarified Yeast Broth
Single-Chain Antibody Fragment
Protein Partitioning
Protein Partitioning
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/30245
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/30245
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purificationPellegrini Malpiedi, LucianaNerli, Bibiana BeatrizAbdalla, Dulcinéia Saes Parrade Alcantara Pessoa, PedroPessoa, Adalberto JrAqueous Micellar Two-Phase SystemCloud PointClarified Yeast BrothSingle-Chain Antibody FragmentProtein PartitioningProtein Partitioninghttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1The main goal of this work was to evaluate the feasibility of using aqueous micellar two-phase systems (AMTPS) of Triton X-114 to purify a single-chain antibody fragment (scFv) directly from yeast fermentation supernatant. The coexistence curves of aqueous micellar two-phase systems, highly loaded (up to 90% wt/wt) with the biological feedstock, were determined. Besides, the effect of several additives such as inorganic salts and affinity ligands on the phase separation behavior was investigated. The obtained coexistence curves demonstrated that the assayed surfactant/yeast broth solutions were able to separate into two phases at temperatures lower than 24 C. This information was then utilized to assay scFv partitioning and purification. In general, proteins present in the yeast broth, including the scFv, partitioned to the top, micelle-depleted phase due to its hydrophilic character. When affinity ligands were used, an opposite behavior was observed for scFv due to the uneven partitioning of ligands toward the micellarrich phase. The best purification performances were attained for the system consisting of 4% wt/wt of Triton X-114, 60% wt/wt of yeast fermentation supernatant and the synthetic ligand Fabsorbent™ F1P HF, with a recovery percentage of 88% and a purification factor of 2. These results demonstrate the potential applicability of these systems for designing early steps for scFv purification directly from yeast broth. New perspectives are now open for the use of this methodology for recombinant antibody fragments purificationFil: Pellegrini Malpiedi, Luciana. Universidade de Sao Paulo; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Quimicos Rosario. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Instituto de Procesos Biotecnológicos y Quimicos Rosario; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Quimicos Rosario. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Instituto de Procesos Biotecnológicos y Quimicos Rosario; ArgentinaFil: Abdalla, Dulcinéia Saes Parra. Universidade de Sao Paulo; BrasilFil: de Alcantara Pessoa, Pedro. Universidade de Sao Paulo; BrasilFil: Pessoa, Adalberto Jr. Universidade de Sao Paulo; BrasilElsevier Science2014-06info: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/11336/30245Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Abdalla, Dulcinéia Saes Parra; de Alcantara Pessoa, Pedro; Pessoa, Adalberto Jr; Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification; Elsevier Science; Separation and Purification Technology; 132; 6-2014; 295-3011383-5866CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1383586614003311info:eu-repo/semantics/altIdentifier/doi/10.1016/j.seppur.2014.05.045info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T10:11:31Zoai:ri.conicet.gov.ar:11336/30245instacron: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 10:11:32.075CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
title Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
spellingShingle Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
Pellegrini Malpiedi, Luciana
Aqueous Micellar Two-Phase System
Cloud Point
Clarified Yeast Broth
Single-Chain Antibody Fragment
Protein Partitioning
Protein Partitioning
title_short Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
title_full Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
title_fullStr Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
title_full_unstemmed Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
title_sort Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification
dc.creator.none.fl_str_mv Pellegrini Malpiedi, Luciana
Nerli, Bibiana Beatriz
Abdalla, Dulcinéia Saes Parra
de Alcantara Pessoa, Pedro
Pessoa, Adalberto Jr
author Pellegrini Malpiedi, Luciana
author_facet Pellegrini Malpiedi, Luciana
Nerli, Bibiana Beatriz
Abdalla, Dulcinéia Saes Parra
de Alcantara Pessoa, Pedro
Pessoa, Adalberto Jr
author_role author
author2 Nerli, Bibiana Beatriz
Abdalla, Dulcinéia Saes Parra
de Alcantara Pessoa, Pedro
Pessoa, Adalberto Jr
author2_role author
author
author
author
dc.subject.none.fl_str_mv Aqueous Micellar Two-Phase System
Cloud Point
Clarified Yeast Broth
Single-Chain Antibody Fragment
Protein Partitioning
Protein Partitioning
topic Aqueous Micellar Two-Phase System
Cloud Point
Clarified Yeast Broth
Single-Chain Antibody Fragment
Protein Partitioning
Protein Partitioning
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The main goal of this work was to evaluate the feasibility of using aqueous micellar two-phase systems (AMTPS) of Triton X-114 to purify a single-chain antibody fragment (scFv) directly from yeast fermentation supernatant. The coexistence curves of aqueous micellar two-phase systems, highly loaded (up to 90% wt/wt) with the biological feedstock, were determined. Besides, the effect of several additives such as inorganic salts and affinity ligands on the phase separation behavior was investigated. The obtained coexistence curves demonstrated that the assayed surfactant/yeast broth solutions were able to separate into two phases at temperatures lower than 24 C. This information was then utilized to assay scFv partitioning and purification. In general, proteins present in the yeast broth, including the scFv, partitioned to the top, micelle-depleted phase due to its hydrophilic character. When affinity ligands were used, an opposite behavior was observed for scFv due to the uneven partitioning of ligands toward the micellarrich phase. The best purification performances were attained for the system consisting of 4% wt/wt of Triton X-114, 60% wt/wt of yeast fermentation supernatant and the synthetic ligand Fabsorbent™ F1P HF, with a recovery percentage of 88% and a purification factor of 2. These results demonstrate the potential applicability of these systems for designing early steps for scFv purification directly from yeast broth. New perspectives are now open for the use of this methodology for recombinant antibody fragments purification
Fil: Pellegrini Malpiedi, Luciana. Universidade de Sao Paulo; Brasil. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Quimicos Rosario. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Instituto de Procesos Biotecnológicos y Quimicos Rosario; Argentina
Fil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Rosario. Instituto de Procesos Biotecnológicos y Quimicos Rosario. Universidad Nacional de Rosario. Facultad de Cs.bioquímicas y Farmaceuticas. Instituto de Procesos Biotecnológicos y Quimicos Rosario; Argentina
Fil: Abdalla, Dulcinéia Saes Parra. Universidade de Sao Paulo; Brasil
Fil: de Alcantara Pessoa, Pedro. Universidade de Sao Paulo; Brasil
Fil: Pessoa, Adalberto Jr. Universidade de Sao Paulo; Brasil
description The main goal of this work was to evaluate the feasibility of using aqueous micellar two-phase systems (AMTPS) of Triton X-114 to purify a single-chain antibody fragment (scFv) directly from yeast fermentation supernatant. The coexistence curves of aqueous micellar two-phase systems, highly loaded (up to 90% wt/wt) with the biological feedstock, were determined. Besides, the effect of several additives such as inorganic salts and affinity ligands on the phase separation behavior was investigated. The obtained coexistence curves demonstrated that the assayed surfactant/yeast broth solutions were able to separate into two phases at temperatures lower than 24 C. This information was then utilized to assay scFv partitioning and purification. In general, proteins present in the yeast broth, including the scFv, partitioned to the top, micelle-depleted phase due to its hydrophilic character. When affinity ligands were used, an opposite behavior was observed for scFv due to the uneven partitioning of ligands toward the micellarrich phase. The best purification performances were attained for the system consisting of 4% wt/wt of Triton X-114, 60% wt/wt of yeast fermentation supernatant and the synthetic ligand Fabsorbent™ F1P HF, with a recovery percentage of 88% and a purification factor of 2. These results demonstrate the potential applicability of these systems for designing early steps for scFv purification directly from yeast broth. New perspectives are now open for the use of this methodology for recombinant antibody fragments purification
publishDate 2014
dc.date.none.fl_str_mv 2014-06
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/11336/30245
Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Abdalla, Dulcinéia Saes Parra; de Alcantara Pessoa, Pedro; Pessoa, Adalberto Jr; Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification; Elsevier Science; Separation and Purification Technology; 132; 6-2014; 295-301
1383-5866
CONICET Digital
CONICET
url http://hdl.handle.net/11336/30245
identifier_str_mv Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Abdalla, Dulcinéia Saes Parra; de Alcantara Pessoa, Pedro; Pessoa, Adalberto Jr; Aqueous micellar systems containing Triton X-114 and Pichia pastoris fermentation supernatant: A novel alternative for single chain-antibody fragment purification; Elsevier Science; Separation and Purification Technology; 132; 6-2014; 295-301
1383-5866
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.sciencedirect.com/science/article/pii/S1383586614003311
info:eu-repo/semantics/altIdentifier/doi/10.1016/j.seppur.2014.05.045
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
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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