Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX
Autor(a) principal: | |
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Data de Publicação: | 2017 |
Outros Autores: | , , |
Tipo de documento: | Artigo |
Idioma: | eng |
Título da fonte: | Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
Texto Completo: | http://hdl.handle.net/10198/20641 |
Resumo: | A DNA aptamer with affinity and specificity for human osteopontin (OPN), a potential breast cancer biomarker, was selected using the SELEX process, considering its homology rate and the stability of its secondary structures. This aptamer exhibited a satisfactory affinity towards OPN, showing dissociation constants lower than 2.5 nM. It was further used to develop a simple, label-free electrochemical aptasensor against OPN. The aptasensor showed good sensitivity towards OPN in standard solutions, being the square wave voltammetry (SWV), compared to the cyclic voltammetry, the most sensitive technique with detection and quantification limits of 1.4 ± 0.4 nM and 4.2 ± 1.1 nM, respectively. It showed good reproducibility and acceptable selectivity, exhibiting low signal interferences from other proteins, as thrombin, with 2.6-10 times lower current signals-off than for OPN. The aptasensor also successfully detected OPN in spiked synthetic human plasma. Using SWV, detection and quantification limits (1.3 ± 0.1 and 3.9 ± 0.4 nM) within the OPN plasma levels reported for patients with breast cancer (0.4-4.5 nM) or with metastatic or recurrent breast cancer (0.9-8.4 nM) were found. Moreover, preliminary assays, using a sample of human plasma, showed that the aptasensor and the standard ELISA method quantified similar OPN levels (2.2 ± 0.7 and 1.7 ± 0.1 nM, respectively). Thus, our aptasensor coupled with SWV represents a promising alternative for the detection of relevant breast cancer biomarkers. |
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Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEXBreast neoplasmsGoldHumansOsteopontinReproducibility of resultsAptamersBiosensing techniquesElectrochemical techniquesA DNA aptamer with affinity and specificity for human osteopontin (OPN), a potential breast cancer biomarker, was selected using the SELEX process, considering its homology rate and the stability of its secondary structures. This aptamer exhibited a satisfactory affinity towards OPN, showing dissociation constants lower than 2.5 nM. It was further used to develop a simple, label-free electrochemical aptasensor against OPN. The aptasensor showed good sensitivity towards OPN in standard solutions, being the square wave voltammetry (SWV), compared to the cyclic voltammetry, the most sensitive technique with detection and quantification limits of 1.4 ± 0.4 nM and 4.2 ± 1.1 nM, respectively. It showed good reproducibility and acceptable selectivity, exhibiting low signal interferences from other proteins, as thrombin, with 2.6-10 times lower current signals-off than for OPN. The aptasensor also successfully detected OPN in spiked synthetic human plasma. Using SWV, detection and quantification limits (1.3 ± 0.1 and 3.9 ± 0.4 nM) within the OPN plasma levels reported for patients with breast cancer (0.4-4.5 nM) or with metastatic or recurrent breast cancer (0.9-8.4 nM) were found. Moreover, preliminary assays, using a sample of human plasma, showed that the aptasensor and the standard ELISA method quantified similar OPN levels (2.2 ± 0.7 and 1.7 ± 0.1 nM, respectively). Thus, our aptasensor coupled with SWV represents a promising alternative for the detection of relevant breast cancer biomarkers.The authors acknowledge the financial support from the Strategic funding of UID/BIO/04469/2013 unit and COMPETE 2020 (POCI-01-0145-FEDER-006684), and from project BioTecNorte (project number NORTE-01-0145-FEDER-000004). This work was also financially supported by Project POCI-01e0145-FEDER-006984 e Associate Laboratory LSRE-LCM and by Project UID/QUI/00616/ 2013 e CQ-VR both funded by FEDER - Fundo Europeu de Desenvolvimento Regional through COMPETE2020 - Programa Operacional Competitividade e Internacionalização (POCI) e and by national funds through FCT - Fundaç~ao para a Ciência e a Tecnologia, Portugal. S. Meirinho also acknowledges the research grant provided by Project UID/EQU/50020/2013.Biblioteca Digital do IPBMeirinho, Sofia G.Dias, L.G.Peres, António M.Rodrigues, Lígia R.2020-02-21T15:49:13Z20172017-01-01T00:00:00Zinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/articleapplication/pdfhttp://hdl.handle.net/10198/20641engMeirinho, Sofia G.; Dias, L.G.; Peres, António M.; Rodrigues, Lígia R. (2017). Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX. Analytica Chimica Acta. ISSN 0003-2670. 987, p. 25-370003-267010.1016/j.aca.2017.07.071info: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:RCAAP2023-11-21T10:46:51Zoai:bibliotecadigital.ipb.pt:10198/20641Portal AgregadorONGhttps://www.rcaap.pt/oai/openaireopendoar:71602024-03-19T23:11:32.820168Repositório 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 |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
title |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
spellingShingle |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX Meirinho, Sofia G. Breast neoplasms Gold Humans Osteopontin Reproducibility of results Aptamers Biosensing techniques Electrochemical techniques |
title_short |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
title_full |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
title_fullStr |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
title_full_unstemmed |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
title_sort |
Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX |
author |
Meirinho, Sofia G. |
author_facet |
Meirinho, Sofia G. Dias, L.G. Peres, António M. Rodrigues, Lígia R. |
author_role |
author |
author2 |
Dias, L.G. Peres, António M. Rodrigues, Lígia R. |
author2_role |
author author author |
dc.contributor.none.fl_str_mv |
Biblioteca Digital do IPB |
dc.contributor.author.fl_str_mv |
Meirinho, Sofia G. Dias, L.G. Peres, António M. Rodrigues, Lígia R. |
dc.subject.por.fl_str_mv |
Breast neoplasms Gold Humans Osteopontin Reproducibility of results Aptamers Biosensing techniques Electrochemical techniques |
topic |
Breast neoplasms Gold Humans Osteopontin Reproducibility of results Aptamers Biosensing techniques Electrochemical techniques |
description |
A DNA aptamer with affinity and specificity for human osteopontin (OPN), a potential breast cancer biomarker, was selected using the SELEX process, considering its homology rate and the stability of its secondary structures. This aptamer exhibited a satisfactory affinity towards OPN, showing dissociation constants lower than 2.5 nM. It was further used to develop a simple, label-free electrochemical aptasensor against OPN. The aptasensor showed good sensitivity towards OPN in standard solutions, being the square wave voltammetry (SWV), compared to the cyclic voltammetry, the most sensitive technique with detection and quantification limits of 1.4 ± 0.4 nM and 4.2 ± 1.1 nM, respectively. It showed good reproducibility and acceptable selectivity, exhibiting low signal interferences from other proteins, as thrombin, with 2.6-10 times lower current signals-off than for OPN. The aptasensor also successfully detected OPN in spiked synthetic human plasma. Using SWV, detection and quantification limits (1.3 ± 0.1 and 3.9 ± 0.4 nM) within the OPN plasma levels reported for patients with breast cancer (0.4-4.5 nM) or with metastatic or recurrent breast cancer (0.9-8.4 nM) were found. Moreover, preliminary assays, using a sample of human plasma, showed that the aptasensor and the standard ELISA method quantified similar OPN levels (2.2 ± 0.7 and 1.7 ± 0.1 nM, respectively). Thus, our aptasensor coupled with SWV represents a promising alternative for the detection of relevant breast cancer biomarkers. |
publishDate |
2017 |
dc.date.none.fl_str_mv |
2017 2017-01-01T00:00:00Z 2020-02-21T15:49:13Z |
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://hdl.handle.net/10198/20641 |
url |
http://hdl.handle.net/10198/20641 |
dc.language.iso.fl_str_mv |
eng |
language |
eng |
dc.relation.none.fl_str_mv |
Meirinho, Sofia G.; Dias, L.G.; Peres, António M.; Rodrigues, Lígia R. (2017). Electrochemical aptasensor for human osteopontin detection using a DNA aptamer selected by SELEX. Analytica Chimica Acta. ISSN 0003-2670. 987, p. 25-37 0003-2670 10.1016/j.aca.2017.07.071 |
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.source.none.fl_str_mv |
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Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) |
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Repositório Científico de Acesso Aberto de Portugal (Repositórios Cientìficos) - Agência para a Sociedade do Conhecimento (UMIC) - FCT - Sociedade da Informação |
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