Bare silica as an alternative matrix for affinity purification/immobilization of His-tagged proteins

Detalhes bibliográficos
Autor(a) principal: Freitas, Ana Isabel
Data de Publicação: 2022
Outros Autores: Domingues, Lucília, Aguiar, Tatiana Quinta
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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spelling 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
instname_str Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação
instacron_str RCAAP
institution 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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