Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity
Autor(a) principal: | |
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Data de Publicação: | 2021 |
Outros Autores: | , , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Institucional da UNESP |
Texto Completo: | http://dx.doi.org/10.1016/j.gce.2021.08.001 http://hdl.handle.net/11449/223399 |
Resumo: | Fluorescent Proteins (FP) can be applied as biomarkers and biosensors in the industrial and medical fields, but their large-scale use, especially for new industrial applications, is limited due to their low stability. Hence, the discovery of additives capable of preserving the activity of FP at room temperature and under stress conditions can help to expand and facilitate their commercial use. With this goal, we evaluated the application of 1-alkyl-3-methylimidazolium chloride-based ([Cnmim]Cl) ionic liquids (ILs) as additives to preserve the activity of Enhanced Green Fluorescent Protein (EGFP) at different storage times and under unfavorable conditions. All [Cnmim]Cl solutions (at 0.100 mol L−1) were able to preserve EGFP fluorescence for longer than the phosphate-saline buffer (PBS) and NaCl solutions, increasing its fluorescence manifestation from 1 to 3 months. [Cnmim]Cl with shorter to medium cationic alkyl chains were the most effective in preserving EGFP fluorescence. [Cnmim]Cl also protected EGFP activity in the presence of the surfactant SDS, the acid guanidine hydrochloride, and H2O2. Therefore, [Cnmim]Cl can be added to aqueous solutions to preserve EGFP fluorescence activity at room temperature for longer storage times and to reduce the negative impact of denaturing agents on EGFP. Therefore, there is a massive potential for the application of ILs as additives to preserve FP in the long-term without refrigeration and under unfavorable conditions, and this is fundamental to enable expansion of FP in industrial and commercial applications. |
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Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activityFluorescenceGreen Fluorescent ProteinIonic liquidsPreservativesProtein stabilityFluorescent Proteins (FP) can be applied as biomarkers and biosensors in the industrial and medical fields, but their large-scale use, especially for new industrial applications, is limited due to their low stability. Hence, the discovery of additives capable of preserving the activity of FP at room temperature and under stress conditions can help to expand and facilitate their commercial use. With this goal, we evaluated the application of 1-alkyl-3-methylimidazolium chloride-based ([Cnmim]Cl) ionic liquids (ILs) as additives to preserve the activity of Enhanced Green Fluorescent Protein (EGFP) at different storage times and under unfavorable conditions. All [Cnmim]Cl solutions (at 0.100 mol L−1) were able to preserve EGFP fluorescence for longer than the phosphate-saline buffer (PBS) and NaCl solutions, increasing its fluorescence manifestation from 1 to 3 months. [Cnmim]Cl with shorter to medium cationic alkyl chains were the most effective in preserving EGFP fluorescence. [Cnmim]Cl also protected EGFP activity in the presence of the surfactant SDS, the acid guanidine hydrochloride, and H2O2. Therefore, [Cnmim]Cl can be added to aqueous solutions to preserve EGFP fluorescence activity at room temperature for longer storage times and to reduce the negative impact of denaturing agents on EGFP. Therefore, there is a massive potential for the application of ILs as additives to preserve FP in the long-term without refrigeration and under unfavorable conditions, and this is fundamental to enable expansion of FP in industrial and commercial applications.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)ASCRS Research FoundationCoordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)Fundação para a Ciência e a TecnologiaDepartment of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP), Rodovia Araraquara-Jaú/Km 01School of Science College of Science Engineering and Health RMIT University, 124 La Trobe StreetSchool of Pharmaceutical Sciences São Paulo University (USP), Av. Prof. Lineu Prestes, no. 580, B16, 05508-000, Cidade de UniversitáriaAustralian Synchrotron, 800 Blackburn RoadUniv Coimbra CIEPQPF Department of Chemical Engineering, Rua Sílvio Lima, Pólo II – Pinhal de MarrocosDepartment of Engineering of Bioprocesses and Biotechnology School of Pharmaceutical Sciences São Paulo State University (UNESP), Rodovia Araraquara-Jaú/Km 01CAPES: 001FAPESP: 2014/16424–7FAPESP: 2014/19793–3FAPESP: 2016/07529–5FAPESP: 2018/01858–2FAPESP: 2018/06576–5FAPESP: 2018/20833–0FAPESP: 2018/25511–1FAPESP: 2018/50009–8FAPESP: 2020/14144–8Fundação para a Ciência e a Tecnologia: UIDB/EQU/00102/2020Fundação para a Ciência e a Tecnologia: UIDP/EQU/00102/2020Universidade Estadual Paulista (UNESP)RMIT UniversityUniversidade de São Paulo (USP)Australian SynchrotronCIEPQPFVeríssimo, Nathalia V. [UNESP]Saponi, Carolina F. [UNESP]Ryan, Timothy M.Greaves, Tamar L.Pereira, Jorge F.B. [UNESP]2022-04-28T19:50:21Z2022-04-28T19:50:21Z2021-12-01info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article412-422http://dx.doi.org/10.1016/j.gce.2021.08.001Green Chemical Engineering, v. 2, n. 4, p. 412-422, 2021.2666-95282096-9147http://hdl.handle.net/11449/22339910.1016/j.gce.2021.08.0012-s2.0-85123982467Scopusreponame:Repositório Institucional da UNESPinstname:Universidade Estadual Paulista (UNESP)instacron:UNESPengGreen Chemical Engineeringinfo:eu-repo/semantics/openAccess2022-04-28T19:50:21Zoai:repositorio.unesp.br:11449/223399Repositório InstitucionalPUBhttp://repositorio.unesp.br/oai/requestopendoar:29462024-08-05T22:25:37.110148Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP)false |
dc.title.none.fl_str_mv |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
title |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
spellingShingle |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity Veríssimo, Nathalia V. [UNESP] Fluorescence Green Fluorescent Protein Ionic liquids Preservatives Protein stability |
title_short |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
title_full |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
title_fullStr |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
title_full_unstemmed |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
title_sort |
Imidazolium-based ionic liquids as additives to preserve the Enhanced Green Fluorescent Protein fluorescent activity |
author |
Veríssimo, Nathalia V. [UNESP] |
author_facet |
Veríssimo, Nathalia V. [UNESP] Saponi, Carolina F. [UNESP] Ryan, Timothy M. Greaves, Tamar L. Pereira, Jorge F.B. [UNESP] |
author_role |
author |
author2 |
Saponi, Carolina F. [UNESP] Ryan, Timothy M. Greaves, Tamar L. Pereira, Jorge F.B. [UNESP] |
author2_role |
author author author author |
dc.contributor.none.fl_str_mv |
Universidade Estadual Paulista (UNESP) RMIT University Universidade de São Paulo (USP) Australian Synchrotron CIEPQPF |
dc.contributor.author.fl_str_mv |
Veríssimo, Nathalia V. [UNESP] Saponi, Carolina F. [UNESP] Ryan, Timothy M. Greaves, Tamar L. Pereira, Jorge F.B. [UNESP] |
dc.subject.por.fl_str_mv |
Fluorescence Green Fluorescent Protein Ionic liquids Preservatives Protein stability |
topic |
Fluorescence Green Fluorescent Protein Ionic liquids Preservatives Protein stability |
description |
Fluorescent Proteins (FP) can be applied as biomarkers and biosensors in the industrial and medical fields, but their large-scale use, especially for new industrial applications, is limited due to their low stability. Hence, the discovery of additives capable of preserving the activity of FP at room temperature and under stress conditions can help to expand and facilitate their commercial use. With this goal, we evaluated the application of 1-alkyl-3-methylimidazolium chloride-based ([Cnmim]Cl) ionic liquids (ILs) as additives to preserve the activity of Enhanced Green Fluorescent Protein (EGFP) at different storage times and under unfavorable conditions. All [Cnmim]Cl solutions (at 0.100 mol L−1) were able to preserve EGFP fluorescence for longer than the phosphate-saline buffer (PBS) and NaCl solutions, increasing its fluorescence manifestation from 1 to 3 months. [Cnmim]Cl with shorter to medium cationic alkyl chains were the most effective in preserving EGFP fluorescence. [Cnmim]Cl also protected EGFP activity in the presence of the surfactant SDS, the acid guanidine hydrochloride, and H2O2. Therefore, [Cnmim]Cl can be added to aqueous solutions to preserve EGFP fluorescence activity at room temperature for longer storage times and to reduce the negative impact of denaturing agents on EGFP. Therefore, there is a massive potential for the application of ILs as additives to preserve FP in the long-term without refrigeration and under unfavorable conditions, and this is fundamental to enable expansion of FP in industrial and commercial applications. |
publishDate |
2021 |
dc.date.none.fl_str_mv |
2021-12-01 2022-04-28T19:50:21Z 2022-04-28T19:50:21Z |
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 |
http://dx.doi.org/10.1016/j.gce.2021.08.001 Green Chemical Engineering, v. 2, n. 4, p. 412-422, 2021. 2666-9528 2096-9147 http://hdl.handle.net/11449/223399 10.1016/j.gce.2021.08.001 2-s2.0-85123982467 |
url |
http://dx.doi.org/10.1016/j.gce.2021.08.001 http://hdl.handle.net/11449/223399 |
identifier_str_mv |
Green Chemical Engineering, v. 2, n. 4, p. 412-422, 2021. 2666-9528 2096-9147 10.1016/j.gce.2021.08.001 2-s2.0-85123982467 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Green Chemical Engineering |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.format.none.fl_str_mv |
412-422 |
dc.source.none.fl_str_mv |
Scopus reponame:Repositório Institucional da UNESP instname:Universidade Estadual Paulista (UNESP) instacron:UNESP |
instname_str |
Universidade Estadual Paulista (UNESP) |
instacron_str |
UNESP |
institution |
UNESP |
reponame_str |
Repositório Institucional da UNESP |
collection |
Repositório Institucional da UNESP |
repository.name.fl_str_mv |
Repositório Institucional da UNESP - Universidade Estadual Paulista (UNESP) |
repository.mail.fl_str_mv |
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1808129426311872512 |