Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems

Autores
Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Taqueda, Maria, E. S.; Abdalla, Dulcineia S. P.; Pessoa Jr., Adalberto
Año de publicación
2015
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
In this work, the purification of a single-chain variable fragment (scFv) of an antibody by using liquid–liquid extraction in aqueous micellar two-phase systems was optimized by means of central composite design. Protein partitioning assays were performed by using the selected system composition in previous works: Triton X-114 at 4% wt/wt, yeast fermentation supernatant at 60% wt/wt, McIlvaine buffer pH 7.00. The other system component concentrations, Cibacron Blue F3GA (CB), Fabsorbent™ F1P HF (HF) and NaCl, were selected as independent variables. ScFv recovery percentage (%R) and purification factor (PF) were selected as the responses. According to the optimization process both, scFv recovery percentage and purification factor were favored with the addition of HF and NaCl in a range of concentrations around the central point of the second central composite design (HF 0.0120% w/w, CB 0.0200% w/w, NaCl 0.200% w/w). These experimental conditions allowed the concentration and pre-purification of scFv in the micelle-rich bottom phase of the systems with a recovery percentage superior to 88% and a purification factor of approximately 3.5. These results improved the previously presented works and demonstrated the convenience of using aqueous micellar two-phase systems as a first step in the purification of scFv molecules.
Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Taqueda, Maria, E. S.. Universidade de Sao Paulo; Brasil
Fil: Abdalla, Dulcineia S. P.. Universidade de Sao Paulo; Brasil
Fil: Pessoa Jr., Adalberto. Universidade de Sao Paulo; Brasil
Materia
Antibody Fragments
Aqueous Micellar Two-Phase Systems
Central Composite Design
Surface Response
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:11336/13402
Acceder
id CONICETDig_32444340698d7a41921d58503513d84f
oai_identifier_str oai:ri.conicet.gov.ar:11336/13402
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systemsPellegrini Malpiedi, LucianaNerli, Bibiana BeatrizTaqueda, Maria, E. S.Abdalla, Dulcineia S. P.Pessoa Jr., AdalbertoAntibody FragmentsAqueous Micellar Two-Phase SystemsCentral Composite DesignSurface Responsehttps://purl.org/becyt/ford/2.11https://purl.org/becyt/ford/2In this work, the purification of a single-chain variable fragment (scFv) of an antibody by using liquid–liquid extraction in aqueous micellar two-phase systems was optimized by means of central composite design. Protein partitioning assays were performed by using the selected system composition in previous works: Triton X-114 at 4% wt/wt, yeast fermentation supernatant at 60% wt/wt, McIlvaine buffer pH 7.00. The other system component concentrations, Cibacron Blue F3GA (CB), Fabsorbent™ F1P HF (HF) and NaCl, were selected as independent variables. ScFv recovery percentage (%R) and purification factor (PF) were selected as the responses. According to the optimization process both, scFv recovery percentage and purification factor were favored with the addition of HF and NaCl in a range of concentrations around the central point of the second central composite design (HF 0.0120% w/w, CB 0.0200% w/w, NaCl 0.200% w/w). These experimental conditions allowed the concentration and pre-purification of scFv in the micelle-rich bottom phase of the systems with a recovery percentage superior to 88% and a purification factor of approximately 3.5. These results improved the previously presented works and demonstrated the convenience of using aqueous micellar two-phase systems as a first step in the purification of scFv molecules.Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario; ArgentinaFil: Taqueda, Maria, E. S.. Universidade de Sao Paulo; BrasilFil: Abdalla, Dulcineia S. P.. Universidade de Sao Paulo; BrasilFil: Pessoa Jr., Adalberto. Universidade de Sao Paulo; BrasilElsevier Inc2015-07info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/13402Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Taqueda, Maria, E. S.; Abdalla, Dulcineia S. P.; Pessoa Jr., Adalberto; Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems; Elsevier Inc; Protein Expression And Purification; 111; 7-2015; 53-601046-5928enginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2015.03.014info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592815000716info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T14:22:19Zoai:ri.conicet.gov.ar:11336/13402instacron: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:34982025-10-15 14:22:20.178CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
title Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
spellingShingle Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
Pellegrini Malpiedi, Luciana
Antibody Fragments
Aqueous Micellar Two-Phase Systems
Central Composite Design
Surface Response
title_short Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
title_full Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
title_fullStr Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
title_full_unstemmed Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
title_sort Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems
dc.creator.none.fl_str_mv Pellegrini Malpiedi, Luciana
Nerli, Bibiana Beatriz
Taqueda, Maria, E. S.
Abdalla, Dulcineia S. P.
Pessoa Jr., Adalberto
author Pellegrini Malpiedi, Luciana
author_facet Pellegrini Malpiedi, Luciana
Nerli, Bibiana Beatriz
Taqueda, Maria, E. S.
Abdalla, Dulcineia S. P.
Pessoa Jr., Adalberto
author_role author
author2 Nerli, Bibiana Beatriz
Taqueda, Maria, E. S.
Abdalla, Dulcineia S. P.
Pessoa Jr., Adalberto
author2_role author
author
author
author
dc.subject.none.fl_str_mv Antibody Fragments
Aqueous Micellar Two-Phase Systems
Central Composite Design
Surface Response
topic Antibody Fragments
Aqueous Micellar Two-Phase Systems
Central Composite Design
Surface Response
purl_subject.fl_str_mv https://purl.org/becyt/ford/2.11
https://purl.org/becyt/ford/2
dc.description.none.fl_txt_mv In this work, the purification of a single-chain variable fragment (scFv) of an antibody by using liquid–liquid extraction in aqueous micellar two-phase systems was optimized by means of central composite design. Protein partitioning assays were performed by using the selected system composition in previous works: Triton X-114 at 4% wt/wt, yeast fermentation supernatant at 60% wt/wt, McIlvaine buffer pH 7.00. The other system component concentrations, Cibacron Blue F3GA (CB), Fabsorbent™ F1P HF (HF) and NaCl, were selected as independent variables. ScFv recovery percentage (%R) and purification factor (PF) were selected as the responses. According to the optimization process both, scFv recovery percentage and purification factor were favored with the addition of HF and NaCl in a range of concentrations around the central point of the second central composite design (HF 0.0120% w/w, CB 0.0200% w/w, NaCl 0.200% w/w). These experimental conditions allowed the concentration and pre-purification of scFv in the micelle-rich bottom phase of the systems with a recovery percentage superior to 88% and a purification factor of approximately 3.5. These results improved the previously presented works and demonstrated the convenience of using aqueous micellar two-phase systems as a first step in the purification of scFv molecules.
Fil: Pellegrini Malpiedi, Luciana. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Nerli, Bibiana Beatriz. Consejo Nacional de Investigaciones Cientificas y Tecnicas. Centro Cientifico Tecnol.conicet - Rosario. Instituto de Procesos Biotecnologicos y Quimicos Rosario; Argentina. Universidad Nacional de Rosario; Argentina
Fil: Taqueda, Maria, E. S.. Universidade de Sao Paulo; Brasil
Fil: Abdalla, Dulcineia S. P.. Universidade de Sao Paulo; Brasil
Fil: Pessoa Jr., Adalberto. Universidade de Sao Paulo; Brasil
description In this work, the purification of a single-chain variable fragment (scFv) of an antibody by using liquid–liquid extraction in aqueous micellar two-phase systems was optimized by means of central composite design. Protein partitioning assays were performed by using the selected system composition in previous works: Triton X-114 at 4% wt/wt, yeast fermentation supernatant at 60% wt/wt, McIlvaine buffer pH 7.00. The other system component concentrations, Cibacron Blue F3GA (CB), Fabsorbent™ F1P HF (HF) and NaCl, were selected as independent variables. ScFv recovery percentage (%R) and purification factor (PF) were selected as the responses. According to the optimization process both, scFv recovery percentage and purification factor were favored with the addition of HF and NaCl in a range of concentrations around the central point of the second central composite design (HF 0.0120% w/w, CB 0.0200% w/w, NaCl 0.200% w/w). These experimental conditions allowed the concentration and pre-purification of scFv in the micelle-rich bottom phase of the systems with a recovery percentage superior to 88% and a purification factor of approximately 3.5. These results improved the previously presented works and demonstrated the convenience of using aqueous micellar two-phase systems as a first step in the purification of scFv molecules.
publishDate 2015
dc.date.none.fl_str_mv 2015-07
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/11336/13402
Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Taqueda, Maria, E. S.; Abdalla, Dulcineia S. P.; Pessoa Jr., Adalberto; Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems; Elsevier Inc; Protein Expression And Purification; 111; 7-2015; 53-60
1046-5928
url http://hdl.handle.net/11336/13402
identifier_str_mv Pellegrini Malpiedi, Luciana; Nerli, Bibiana Beatriz; Taqueda, Maria, E. S.; Abdalla, Dulcineia S. P.; Pessoa Jr., Adalberto; Optimized extraction of a single-chain variable fragment of antibody by using aqueous micellar two-phase systems; Elsevier Inc; Protein Expression And Purification; 111; 7-2015; 53-60
1046-5928
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.pep.2015.03.014
info:eu-repo/semantics/altIdentifier/url/http://www.sciencedirect.com/science/article/pii/S1046592815000716
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 Elsevier Inc
publisher.none.fl_str_mv Elsevier Inc
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
_version_ 1846082622215684096
score 13.22299

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