Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography
- Autores
- Levin, Gustavo Javier; Mendive, Fernando; Targovnik, Hector Manuel; Cascone, Osvaldo; Miranda, Maria Victoria
- Año de publicación
- 2005
- Idioma
- inglés
- Tipo de recurso
- artículo
- Estado
- versión publicada
- Descripción
- An engineered horseradish peroxidase isozyme C (HRP C) gene was constructed by the addition of a 6xArg fusion tail to 6xHis-HRP C by the PCR strategy. The 6xHis-6xArg-HRP C cDNA was expressed in the Sf9 insect cell line from Spodoptera frugiperda infected with Autographa californica nuclear polyhedrosis virus. The recombinant peroxidase isoelectric point was 9.5 as judged by isoelectric focusing and was purified directly from the culture medium at day-6 post-infection by cation-exchange chromatography or immobilised metal ion-affinity chromatography. While the former technique gave a yield of 98.5% with a purification factor of 130, the latter gave only a 68% yield with a purification factor of 140. Results obtained provide evidence that the poly-Arg tag is more effective than the poly-His tag for peroxidase purification from a baculovirus expression system.
Fil: Levin, Gustavo Javier. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Mendive, Fernando. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Genética y Biología Molecular; Argentina
Fil: Targovnik, Hector Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Genética y Biología Molecular; Argentina
Fil: Cascone, Osvaldo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina
Fil: Miranda, Maria Victoria. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina - Materia
-
FUSION TAIL
IMMOBILISED METAL ION-AFFINITY CHROMATOGRAPHY
ION-EXCHANGE CHROMATOGRAPHY
PEROXIDASE
PURIFICATION - Nivel de accesibilidad
- acceso abierto
- Condiciones de uso
- https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
- Repositorio
- Institución
- Consejo Nacional de Investigaciones Científicas y Técnicas
- OAI Identificador
- oai:ri.conicet.gov.ar:11336/116258
Ver los metadatos del registro completo
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Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatographyLevin, Gustavo JavierMendive, FernandoTargovnik, Hector ManuelCascone, OsvaldoMiranda, Maria VictoriaFUSION TAILIMMOBILISED METAL ION-AFFINITY CHROMATOGRAPHYION-EXCHANGE CHROMATOGRAPHYPEROXIDASEPURIFICATIONhttps://purl.org/becyt/ford/2.11https://purl.org/becyt/ford/2An engineered horseradish peroxidase isozyme C (HRP C) gene was constructed by the addition of a 6xArg fusion tail to 6xHis-HRP C by the PCR strategy. The 6xHis-6xArg-HRP C cDNA was expressed in the Sf9 insect cell line from Spodoptera frugiperda infected with Autographa californica nuclear polyhedrosis virus. The recombinant peroxidase isoelectric point was 9.5 as judged by isoelectric focusing and was purified directly from the culture medium at day-6 post-infection by cation-exchange chromatography or immobilised metal ion-affinity chromatography. While the former technique gave a yield of 98.5% with a purification factor of 130, the latter gave only a 68% yield with a purification factor of 140. Results obtained provide evidence that the poly-Arg tag is more effective than the poly-His tag for peroxidase purification from a baculovirus expression system.Fil: Levin, Gustavo Javier. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Mendive, Fernando. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Genética y Biología Molecular; ArgentinaFil: Targovnik, Hector Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Genética y Biología Molecular; ArgentinaFil: Cascone, Osvaldo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Miranda, Maria Victoria. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaElsevier Science2005-09-10info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/116258Levin, Gustavo Javier; Mendive, Fernando; Targovnik, Hector Manuel; Cascone, Osvaldo; Miranda, Maria Victoria; Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography; Elsevier Science; Journal of Biotechnology; 118; 4; 10-9-2005; 363-3690168-1656CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.jbiotec.2005.05.015info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0168165605002403?via%3Dihubinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-29T10:39:01Zoai:ri.conicet.gov.ar:11336/116258instacron: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 10:39:02.009CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse |
dc.title.none.fl_str_mv |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
title |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
spellingShingle |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography Levin, Gustavo Javier FUSION TAIL IMMOBILISED METAL ION-AFFINITY CHROMATOGRAPHY ION-EXCHANGE CHROMATOGRAPHY PEROXIDASE PURIFICATION |
title_short |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
title_full |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
title_fullStr |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
title_full_unstemmed |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
title_sort |
Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography |
dc.creator.none.fl_str_mv |
Levin, Gustavo Javier Mendive, Fernando Targovnik, Hector Manuel Cascone, Osvaldo Miranda, Maria Victoria |
author |
Levin, Gustavo Javier |
author_facet |
Levin, Gustavo Javier Mendive, Fernando Targovnik, Hector Manuel Cascone, Osvaldo Miranda, Maria Victoria |
author_role |
author |
author2 |
Mendive, Fernando Targovnik, Hector Manuel Cascone, Osvaldo Miranda, Maria Victoria |
author2_role |
author author author author |
dc.subject.none.fl_str_mv |
FUSION TAIL IMMOBILISED METAL ION-AFFINITY CHROMATOGRAPHY ION-EXCHANGE CHROMATOGRAPHY PEROXIDASE PURIFICATION |
topic |
FUSION TAIL IMMOBILISED METAL ION-AFFINITY CHROMATOGRAPHY ION-EXCHANGE CHROMATOGRAPHY PEROXIDASE PURIFICATION |
purl_subject.fl_str_mv |
https://purl.org/becyt/ford/2.11 https://purl.org/becyt/ford/2 |
dc.description.none.fl_txt_mv |
An engineered horseradish peroxidase isozyme C (HRP C) gene was constructed by the addition of a 6xArg fusion tail to 6xHis-HRP C by the PCR strategy. The 6xHis-6xArg-HRP C cDNA was expressed in the Sf9 insect cell line from Spodoptera frugiperda infected with Autographa californica nuclear polyhedrosis virus. The recombinant peroxidase isoelectric point was 9.5 as judged by isoelectric focusing and was purified directly from the culture medium at day-6 post-infection by cation-exchange chromatography or immobilised metal ion-affinity chromatography. While the former technique gave a yield of 98.5% with a purification factor of 130, the latter gave only a 68% yield with a purification factor of 140. Results obtained provide evidence that the poly-Arg tag is more effective than the poly-His tag for peroxidase purification from a baculovirus expression system. Fil: Levin, Gustavo Javier. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Mendive, Fernando. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Genética y Biología Molecular; Argentina Fil: Targovnik, Hector Manuel. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay. Instituto de Inmunología, Genética y Metabolismo. Universidad de Buenos Aires. Facultad de Medicina. Instituto de Inmunología, Genética y Metabolismo; Argentina. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Genética y Biología Molecular; Argentina Fil: Cascone, Osvaldo. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina Fil: Miranda, Maria Victoria. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Microbiología Industrial y Biotecnología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; Argentina |
description |
An engineered horseradish peroxidase isozyme C (HRP C) gene was constructed by the addition of a 6xArg fusion tail to 6xHis-HRP C by the PCR strategy. The 6xHis-6xArg-HRP C cDNA was expressed in the Sf9 insect cell line from Spodoptera frugiperda infected with Autographa californica nuclear polyhedrosis virus. The recombinant peroxidase isoelectric point was 9.5 as judged by isoelectric focusing and was purified directly from the culture medium at day-6 post-infection by cation-exchange chromatography or immobilised metal ion-affinity chromatography. While the former technique gave a yield of 98.5% with a purification factor of 130, the latter gave only a 68% yield with a purification factor of 140. Results obtained provide evidence that the poly-Arg tag is more effective than the poly-His tag for peroxidase purification from a baculovirus expression system. |
publishDate |
2005 |
dc.date.none.fl_str_mv |
2005-09-10 |
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/11336/116258 Levin, Gustavo Javier; Mendive, Fernando; Targovnik, Hector Manuel; Cascone, Osvaldo; Miranda, Maria Victoria; Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography; Elsevier Science; Journal of Biotechnology; 118; 4; 10-9-2005; 363-369 0168-1656 CONICET Digital CONICET |
url |
http://hdl.handle.net/11336/116258 |
identifier_str_mv |
Levin, Gustavo Javier; Mendive, Fernando; Targovnik, Hector Manuel; Cascone, Osvaldo; Miranda, Maria Victoria; Genetically engineered horseradish peroxidase for facilitated purification from baculovirus cultures by cation-exchange chromatography; Elsevier Science; Journal of Biotechnology; 118; 4; 10-9-2005; 363-369 0168-1656 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.jbiotec.2005.05.015 info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0168165605002403?via%3Dihub |
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 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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1844614414565638144 |
score |
13.070432 |