The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system

Autores
Muruaga, Emanuel Javier; Uriza, Paula Jimena; Eckert, Gonzalo Axel Klaus; Pepe, María Victoria; Duarte, Cecilia Magalí; Roset, Mara Sabrina; Briones, Carlos Gabriel
Año de publicación
2022
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
The SLAPTAG is a novel molecular TAG derived from a protein domain present in the sequence of Lactobacillus acidophilus SlpA (SlpA284-444). Proteins from different biological sources, with different molecular weights or biochemical functions, can be fused in frame to the SLAPTAG and efficiently purified by the specific binding to a bacterial-derived chromatographic matrix named here Bio-Matrix (BM). To set an optimized protocol for the SLAPTAG-based affinity chromatography (SAC) different binding and elution conditions were evaluated. The binding equilibrium between SLAPTAG and BM was reached after a few minutes at 4°C, being the apparent dissociation constant (KD) of 4.3 uM, a value similar to the one determined for other S-layer proteins and their respective bacterial cell-walls. A reporter protein was generated (H6-GFP-SLAPTAG) to compare the efficiency of SAC against a commercial system based on a Ni2+-charged agarose matrix, observing no differences in the H6-GFP-SLAPTAG purification performance. The stability and reusability of the BM were evaluated, and it was determined that the matrix was stable for more than a year, being possible to reuse it five times without a significant loss in the efficiency for protein purification. Alternatively, we explored the recovery of bound SLAP-tagged proteins by proteolysis using the SLAPASE (a SLAP-tagged version of the HRV-3c protease) that released a tag-less GFP (SLAPTAG-less). Additionally, iron nanoparticles were linked to the BM and the resulting BMmag was successfully adapted for a magnetic SAC, a technique that can be potentially applied for high-throughput-out protein production and purification.
Fil: Muruaga, Emanuel Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Uriza, Paula Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Eckert, Gonzalo Axel Klaus. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Pepe, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Duarte, Cecilia Magalí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Roset, Mara Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Briones, Carlos Gabriel. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; Argentina
Materia
AFFINITY CHROMATOGRAPHY
SLAP
RECOMBINANT PROTEINS
BACILLUS
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:11336/215848
Acceder
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network_name_str CONICET Digital (CONICET)
spelling The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography systemMuruaga, Emanuel JavierUriza, Paula JimenaEckert, Gonzalo Axel KlausPepe, María VictoriaDuarte, Cecilia MagalíRoset, Mara SabrinaBriones, Carlos GabrielAFFINITY CHROMATOGRAPHYSLAPRECOMBINANT PROTEINSBACILLUShttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1The SLAPTAG is a novel molecular TAG derived from a protein domain present in the sequence of Lactobacillus acidophilus SlpA (SlpA284-444). Proteins from different biological sources, with different molecular weights or biochemical functions, can be fused in frame to the SLAPTAG and efficiently purified by the specific binding to a bacterial-derived chromatographic matrix named here Bio-Matrix (BM). To set an optimized protocol for the SLAPTAG-based affinity chromatography (SAC) different binding and elution conditions were evaluated. The binding equilibrium between SLAPTAG and BM was reached after a few minutes at 4°C, being the apparent dissociation constant (KD) of 4.3 uM, a value similar to the one determined for other S-layer proteins and their respective bacterial cell-walls. A reporter protein was generated (H6-GFP-SLAPTAG) to compare the efficiency of SAC against a commercial system based on a Ni2+-charged agarose matrix, observing no differences in the H6-GFP-SLAPTAG purification performance. The stability and reusability of the BM were evaluated, and it was determined that the matrix was stable for more than a year, being possible to reuse it five times without a significant loss in the efficiency for protein purification. Alternatively, we explored the recovery of bound SLAP-tagged proteins by proteolysis using the SLAPASE (a SLAP-tagged version of the HRV-3c protease) that released a tag-less GFP (SLAPTAG-less). Additionally, iron nanoparticles were linked to the BM and the resulting BMmag was successfully adapted for a magnetic SAC, a technique that can be potentially applied for high-throughput-out protein production and purification.Fil: Muruaga, Emanuel Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Uriza, Paula Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Eckert, Gonzalo Axel Klaus. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Pepe, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Duarte, Cecilia Magalí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Roset, Mara Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; ArgentinaFil: Briones, Carlos Gabriel. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; ArgentinaCold Spring Harbor Laboratory Press2022-12info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/215848Muruaga, Emanuel Javier; Uriza, Paula Jimena; Eckert, Gonzalo Axel Klaus; Pepe, María Victoria; Duarte, Cecilia Magalí; et al.; The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system; Cold Spring Harbor Laboratory Press; BioRXiv; 2023; 12-2022; 1-322692-8205CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2022.12.24.521862v2info:eu-repo/semantics/altIdentifier/doi/10.1101/2022.12.24.521862info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-nd/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-10-15T15:01:02Zoai:ri.conicet.gov.ar:11336/215848instacron: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 15:01:02.468CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
title The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
spellingShingle The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
Muruaga, Emanuel Javier
AFFINITY CHROMATOGRAPHY
SLAP
RECOMBINANT PROTEINS
BACILLUS
title_short The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
title_full The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
title_fullStr The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
title_full_unstemmed The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
title_sort The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system
dc.creator.none.fl_str_mv Muruaga, Emanuel Javier
Uriza, Paula Jimena
Eckert, Gonzalo Axel Klaus
Pepe, María Victoria
Duarte, Cecilia Magalí
Roset, Mara Sabrina
Briones, Carlos Gabriel
author Muruaga, Emanuel Javier
author_facet Muruaga, Emanuel Javier
Uriza, Paula Jimena
Eckert, Gonzalo Axel Klaus
Pepe, María Victoria
Duarte, Cecilia Magalí
Roset, Mara Sabrina
Briones, Carlos Gabriel
author_role author
author2 Uriza, Paula Jimena
Eckert, Gonzalo Axel Klaus
Pepe, María Victoria
Duarte, Cecilia Magalí
Roset, Mara Sabrina
Briones, Carlos Gabriel
author2_role author
author
author
author
author
author
dc.subject.none.fl_str_mv AFFINITY CHROMATOGRAPHY
SLAP
RECOMBINANT PROTEINS
BACILLUS
topic AFFINITY CHROMATOGRAPHY
SLAP
RECOMBINANT PROTEINS
BACILLUS
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv The SLAPTAG is a novel molecular TAG derived from a protein domain present in the sequence of Lactobacillus acidophilus SlpA (SlpA284-444). Proteins from different biological sources, with different molecular weights or biochemical functions, can be fused in frame to the SLAPTAG and efficiently purified by the specific binding to a bacterial-derived chromatographic matrix named here Bio-Matrix (BM). To set an optimized protocol for the SLAPTAG-based affinity chromatography (SAC) different binding and elution conditions were evaluated. The binding equilibrium between SLAPTAG and BM was reached after a few minutes at 4°C, being the apparent dissociation constant (KD) of 4.3 uM, a value similar to the one determined for other S-layer proteins and their respective bacterial cell-walls. A reporter protein was generated (H6-GFP-SLAPTAG) to compare the efficiency of SAC against a commercial system based on a Ni2+-charged agarose matrix, observing no differences in the H6-GFP-SLAPTAG purification performance. The stability and reusability of the BM were evaluated, and it was determined that the matrix was stable for more than a year, being possible to reuse it five times without a significant loss in the efficiency for protein purification. Alternatively, we explored the recovery of bound SLAP-tagged proteins by proteolysis using the SLAPASE (a SLAP-tagged version of the HRV-3c protease) that released a tag-less GFP (SLAPTAG-less). Additionally, iron nanoparticles were linked to the BM and the resulting BMmag was successfully adapted for a magnetic SAC, a technique that can be potentially applied for high-throughput-out protein production and purification.
Fil: Muruaga, Emanuel Javier. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Uriza, Paula Jimena. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Eckert, Gonzalo Axel Klaus. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Pepe, María Victoria. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Duarte, Cecilia Magalí. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Roset, Mara Sabrina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - La Plata. Instituto de Investigaciones Biotecnológicas. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas; Argentina
Fil: Briones, Carlos Gabriel. Universidad Nacional de San Martín. Instituto de Investigaciones Biotecnológicas. - Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Parque Centenario. Instituto de Investigaciones Biotecnológicas; Argentina
description The SLAPTAG is a novel molecular TAG derived from a protein domain present in the sequence of Lactobacillus acidophilus SlpA (SlpA284-444). Proteins from different biological sources, with different molecular weights or biochemical functions, can be fused in frame to the SLAPTAG and efficiently purified by the specific binding to a bacterial-derived chromatographic matrix named here Bio-Matrix (BM). To set an optimized protocol for the SLAPTAG-based affinity chromatography (SAC) different binding and elution conditions were evaluated. The binding equilibrium between SLAPTAG and BM was reached after a few minutes at 4°C, being the apparent dissociation constant (KD) of 4.3 uM, a value similar to the one determined for other S-layer proteins and their respective bacterial cell-walls. A reporter protein was generated (H6-GFP-SLAPTAG) to compare the efficiency of SAC against a commercial system based on a Ni2+-charged agarose matrix, observing no differences in the H6-GFP-SLAPTAG purification performance. The stability and reusability of the BM were evaluated, and it was determined that the matrix was stable for more than a year, being possible to reuse it five times without a significant loss in the efficiency for protein purification. Alternatively, we explored the recovery of bound SLAP-tagged proteins by proteolysis using the SLAPASE (a SLAP-tagged version of the HRV-3c protease) that released a tag-less GFP (SLAPTAG-less). Additionally, iron nanoparticles were linked to the BM and the resulting BMmag was successfully adapted for a magnetic SAC, a technique that can be potentially applied for high-throughput-out protein production and purification.
publishDate 2022
dc.date.none.fl_str_mv 2022-12
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/11336/215848
Muruaga, Emanuel Javier; Uriza, Paula Jimena; Eckert, Gonzalo Axel Klaus; Pepe, María Victoria; Duarte, Cecilia Magalí; et al.; The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system; Cold Spring Harbor Laboratory Press; BioRXiv; 2023; 12-2022; 1-32
2692-8205
CONICET Digital
CONICET
url http://hdl.handle.net/11336/215848
identifier_str_mv Muruaga, Emanuel Javier; Uriza, Paula Jimena; Eckert, Gonzalo Axel Klaus; Pepe, María Victoria; Duarte, Cecilia Magalí; et al.; The SLAPTAG: A new molecular tag adapted for the development of a high-performance, low-cost, affinity chromatography system; Cold Spring Harbor Laboratory Press; BioRXiv; 2023; 12-2022; 1-32
2692-8205
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/url/https://www.biorxiv.org/content/10.1101/2022.12.24.521862v2
info:eu-repo/semantics/altIdentifier/doi/10.1101/2022.12.24.521862
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 Cold Spring Harbor Laboratory Press
publisher.none.fl_str_mv Cold Spring Harbor Laboratory Press
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 13.22299

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