Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins
Autor(a) principal: | |
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Data de Publicação: | 2022 |
Outros Autores: | , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | https://hdl.handle.net/1822/75759 |
Resumo: | Supplementary data to this article can be found online at: https://doi. org/10.1016/j.seppur.2022.120448. |
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Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteinsSilicon dioxideSilica-binding peptidesHis-tagAffinity purification/immobilizationBatch affinity chromatographyAffinity purificationimmobilizationScience & TechnologySupplementary data to this article can be found online at: https://doi. org/10.1016/j.seppur.2022.120448.Green protein purification/immobilization processes based on low-cost, earth-abundant, and eco-friendly affinity matrices are highly desirable. Unmodified silica matrices fit well these demands. Since histidine-rich silica-binding peptides are frequently isolated in biopanning experiments, this work aimed at assessing the viability of using bare silica as an alternative matrix for the purification/immobilization of His-tagged proteins. Adsorption and desorption studies with a purified His6-tagged EGFP shown that binding to bare silica particles of different size and porosity occurred under the conditions tested, and that elution could be accomplished with eco-friendly eluants containing L-arginine/L-lysine. Non-tagged EGFP did not bind to these matrices. Small-scale batch purification schemes using silica gel Davisil grade 643 or 646 as affinity matrices and a Tris-buffered saline eluant containing 0.5 M L-arginine (pH 8.5) allowed purifying His6-EGFP from Escherichia coli lysates with a purity of up to 96 % and a recovery yield of 70 % after just one elution step. EGFP tagged with the silica-binding peptide Car9 was recovered with comparable purity and yield. Other His-tagged proteins could also be purified to similar purity levels. The scale of this batch purification scheme was shown to be extendable. These results demonstrate that unmodified silica matrices can be used to effectively purify His-tagged proteins. Since the recovery of double tagged His6-EGFP-Car9 was only of 30-55 %, the combination of tags revealed to be advantageous for immobilization purposes.This study was supported by the Portuguese Foundation for Science and Technology (FCT) under the scope of the strategic funding of UIDB/04469/2020 unit and Project EcoBioInks4SmartTextiles [PTDC/CTM-TEX/30298/2017–POCI-01-0145-FEDER-030298], co-funded by the European Regional Development Fund (ERDF) through the Competitiveness and Internationalization Operational Programme (COMPETE 2020) of Portugal 2020.info:eu-repo/semantics/publishedVersionElsevierUniversidade do MinhoFreitas, Ana IsabelDomingues, LucíliaAguiar, Tatiana Quinta2022-042022-04-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttps://hdl.handle.net/1822/75759engFreitas, Ana Isabel; Domingues, Lucília; Aguiar, Tatiana Q., Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins. Separation and Purification Technology, 286(120448), 20221383-586610.1016/j.seppur.2022.120448120448https://www.sciencedirect.com/science/article/pii/S1383586622000089info:eu-repo/semantics/openAccessreponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos)instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãoinstacron:RCAAP2023-07-21T12:48:41Zoai:repositorium.sdum.uminho.pt:1822/75759Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T19:46:59.890939Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informaçãofalse |
dc.title.none.fl_str_mv |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
title |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
spellingShingle |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins Freitas, Ana Isabel Silicon dioxide Silica-binding peptides His-tag Affinity purification/immobilization Batch affinity chromatography Affinity purification immobilization Science & Technology |
title_short |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
title_full |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
title_fullStr |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
title_full_unstemmed |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
title_sort |
Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins |
author |
Freitas, Ana Isabel |
author_facet |
Freitas, Ana Isabel Domingues, Lucília Aguiar, Tatiana Quinta |
author_role |
author |
author2 |
Domingues, Lucília Aguiar, Tatiana Quinta |
author2_role |
author author |
dc.contributor.none.fl_str_mv |
Universidade do Minho |
dc.contributor.author.fl_str_mv |
Freitas, Ana Isabel Domingues, Lucília Aguiar, Tatiana Quinta |
dc.subject.por.fl_str_mv |
Silicon dioxide Silica-binding peptides His-tag Affinity purification/immobilization Batch affinity chromatography Affinity purification immobilization Science & Technology |
topic |
Silicon dioxide Silica-binding peptides His-tag Affinity purification/immobilization Batch affinity chromatography Affinity purification immobilization Science & Technology |
description |
Supplementary data to this article can be found online at: https://doi. org/10.1016/j.seppur.2022.120448. |
publishDate |
2022 |
dc.date.none.fl_str_mv |
2022-04 2022-04-01T00:00:00Z |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/article |
format |
article |
status_str |
publishedVersion |
dc.identifier.uri.fl_str_mv |
https://hdl.handle.net/1822/75759 |
url |
https://hdl.handle.net/1822/75759 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Freitas, Ana Isabel; Domingues, Lucília; Aguiar, Tatiana Q., Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins. Separation and Purification Technology, 286(120448), 2022 1383-5866 10.1016/j.seppur.2022.120448 120448 https://www.sciencedirect.com/science/article/pii/S1383586622000089 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
application/pdf |
dc.publisher.none.fl_str_mv |
Elsevier |
publisher.none.fl_str_mv |
Elsevier |
dc.source.none.fl_str_mv |
reponame:Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) instname:Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação instacron:RCAAP |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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RCAAP |
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RCAAP |
reponame_str |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
collection |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
repository.name.fl_str_mv |
Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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1799133041004642304 |