Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro
Autor(a) principal: | |
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Data de Publicação: | 2019 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/11031 |
Resumo: | In the present work, nanostructured films were prepared through the self-assembly techniques for the detection of prostatic specific antigen (PSA), a glycoprotein produced in abnormal levels in case of prostate cancer. Two self-assembly approaches were used: the self-assembled monolayer (SAM) technique for the substrate functionalization and the layer-by-layer (LbL) technique for the building of the sensing unit. The specific antibody to PSA (anti-PSA) was immobilized on the gold nanoparticles surface (AuNP-antiPSA conjugate) for later assembly by the LbL technique. The AuNPs-anti-PSA conjugate was characterized by ultraviolet-visible spectroscopy indicating absorption bands at 520 and 260 nm related to the AuNP plasmons absorption and anti-PSA tryptophan absorption, respectively. The fluorescence spectrum presented a band at 367 nm attributed to the emission of tryptophan fluorescence present in the anti-PSA. The morphology analysis of AuNPs by Transmission Electron Microscopy (MET) provided an estimated average diameter of 3.5 ± 0.9 nm. The films were fabricated on gold (Au) covered glass slides previously cleaned and functionalized with 11-mercaptoundecanoic acid (11-MUA). The LbL films were prepared by the sequential adsorption of nanostructured layers of poly(ethyleneimine) (PEI), poly(vinylsulfonic acid, sodium salt) (PVS) and PEI, on the Au functionalized surface. Cyclic voltammetry (VC) confirmed the adsorption of the layers by the reduction of the current peak of Fe(II)/Fe(III) redox probe. Subsequently, bovine serum albumin (BSA) was used to block the non-specific binding sites. Detection was performed with different concentrations of PSA (0.1, 0.5, 1,0, 4,0, 8,0, 10 and 20 ng mL-1) using Electrochemical Impedance Spectroscopy (EIS). The architecture Au/11-MUA/PEI-PVS-PEI/AuNPs-anti-PSA/BSA/PSA was also characterized by Polarization-Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS) to investigate the interaction between the materials, as well as the interaction between antigen and antibody. The high selectivity of the immunosensor was tested by the exposure of the sensor to a non-specific immunoglobulin G (IgG). The limit of detection (LD) and the range of detection obtained by EIS were 0.17 ng mL-1 and 0.1-20 ng mL-1, respectively. The experiments demonstrated that the LbL technique and the use of AuNPs were suitable for the construction of an immunosensor of low cost, rapid development and high sensitivity. Thus, the fabricated immunoassay exhibits high potential for use in the diagnosis of the early stages of prostate cancer. |
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Camilo, Douglas EleutérioFerreira, Marystelahttp://lattes.cnpq.br/4714185516263478Massumi Miyazaki, Celinahttp://lattes.cnpq.br/2932962321837120http://lattes.cnpq.br/184459611918527950fc25b9-3a47-4a4a-9d23-b936eca9185c2019-02-26T18:16:55Z2019-02-26T18:16:55Z2019-02-15CAMILO, Douglas Eleutério. Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro. 2019. Dissertação (Mestrado em Ciência dos Materiais) – Universidade Federal de São Carlos, Sorocaba, 2019. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11031.https://repositorio.ufscar.br/handle/ufscar/11031In the present work, nanostructured films were prepared through the self-assembly techniques for the detection of prostatic specific antigen (PSA), a glycoprotein produced in abnormal levels in case of prostate cancer. Two self-assembly approaches were used: the self-assembled monolayer (SAM) technique for the substrate functionalization and the layer-by-layer (LbL) technique for the building of the sensing unit. The specific antibody to PSA (anti-PSA) was immobilized on the gold nanoparticles surface (AuNP-antiPSA conjugate) for later assembly by the LbL technique. The AuNPs-anti-PSA conjugate was characterized by ultraviolet-visible spectroscopy indicating absorption bands at 520 and 260 nm related to the AuNP plasmons absorption and anti-PSA tryptophan absorption, respectively. The fluorescence spectrum presented a band at 367 nm attributed to the emission of tryptophan fluorescence present in the anti-PSA. The morphology analysis of AuNPs by Transmission Electron Microscopy (MET) provided an estimated average diameter of 3.5 ± 0.9 nm. The films were fabricated on gold (Au) covered glass slides previously cleaned and functionalized with 11-mercaptoundecanoic acid (11-MUA). The LbL films were prepared by the sequential adsorption of nanostructured layers of poly(ethyleneimine) (PEI), poly(vinylsulfonic acid, sodium salt) (PVS) and PEI, on the Au functionalized surface. Cyclic voltammetry (VC) confirmed the adsorption of the layers by the reduction of the current peak of Fe(II)/Fe(III) redox probe. Subsequently, bovine serum albumin (BSA) was used to block the non-specific binding sites. Detection was performed with different concentrations of PSA (0.1, 0.5, 1,0, 4,0, 8,0, 10 and 20 ng mL-1) using Electrochemical Impedance Spectroscopy (EIS). The architecture Au/11-MUA/PEI-PVS-PEI/AuNPs-anti-PSA/BSA/PSA was also characterized by Polarization-Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS) to investigate the interaction between the materials, as well as the interaction between antigen and antibody. The high selectivity of the immunosensor was tested by the exposure of the sensor to a non-specific immunoglobulin G (IgG). The limit of detection (LD) and the range of detection obtained by EIS were 0.17 ng mL-1 and 0.1-20 ng mL-1, respectively. The experiments demonstrated that the LbL technique and the use of AuNPs were suitable for the construction of an immunosensor of low cost, rapid development and high sensitivity. Thus, the fabricated immunoassay exhibits high potential for use in the diagnosis of the early stages of prostate cancer.Neste trabalho foram produzidos filmes nanoestruturados para a detecção do antígeno prostático específico (PSA), uma glicoproteína produzida em quantidades anormais em caso de câncer de próstata, utilizando-se de técnicas de automontagem: a monocamada automontada (SAM) para a funcionalização do substrato, seguida da camada por camada (LbL) para a fabricação das multicamadas que compõe a unidade sensorial. Cada etapa de adsorção foi monitorada por diferentes técnicas de caracterização. O anticorpo específico do PSA (anti-PSA) foi imobilizado na superfície de nanopartículas de ouro (conjugado AuNPs-antiPSA) para a posterior montagem pela técnica LbL. As AuNPs-anti-PSA foram caracterizadas por espectroscopia no ultravioleta-visível (UV-Vis), que indicou bandas de absorção plasmônica das AuNPs em 520 nm e de absorção do triptofano presente no anti-PSA em 260 nm. Com a espectroscopia de fluorescência verificou-se a banda de emissão de fluorescência do triptofano em 367 nm. A análise da morfologia das AuNPs por microscopia eletrônica de transmissão (MET) forneceu o diâmetro médio estimado de 3,5 ± 0,9 nm. Os filmes foram fabricados sobre lâminas de ouro (Au) previamente limpas e funcionalizadas com ácido 11-mercaptoundecanóico (11-MUA). Os filmes LbL foram preparados com a adsorção de uma tricamada de polieletrólitos composto por poli(etilenoimina) (PEI), poli(ácido vinilsulfônico, sal de sódio) (PVS) e novamente PEI, sobre o Au funcionalizado com 11-MUA, e caracterizados por voltametria cíclica (VC). Em seguida foi imobilizado o conjugado AuNP-antiPSA e por fim, a albumina de soro bovino (BSA) foi utilizada para bloquear os sítios de ligação não específicos. Observou-se a diminuição da corrente nos picos redox de Fe(II)/Fe(III) com a adsorção sucessiva das camadas. A detecção foi realizada com diferentes concentrações de PSA (0,1, 0,5, 1,0, 4,0, 8,0, 10 e 20 ng mL-1) utilizando a espectroscopia de impedância eletroquímica (EIE). A arquitetura descrita, Au/11-MUA/PEI-PVS-PEI/AuNPs-anti-PSA/BSA/PSA, foi também caracterizada por espectroscopia de reflexão-absorção no infravermelho com modulação da polarização (PM-IRRAS) para se identificar cada material, a interação entre eles, bem como a interação entre o antígeno e o anticorpo. A seletividade do imunossensor foi analisada através da exposição à imunoglobulina G (IgG). O limite de detecção (LD) e a faixa analítica obtidos da EIE foiram, respectivamente, de 0,17 ng mL-1 e 0,1-20 ng mL-1. Os experimentos evidenciaram que a técnica LbL e o uso de AuNPs foram adequados para a construção de um imunossensor de baixo custo, rápido desenvolvimento e alta sensibilidade. Sendo assim, o imunossensor fabricado exibe significativo potencial para ser utilizado no diagnóstico do câncer de próstata.Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)1687177porUniversidade Federal de São CarlosCâmpus SorocabaPrograma de Pós-Graduação em Ciência dos Materiais - PPGCM-SoUFSCarFilmes camada por camadaAnticorpo anti-PSAAntígeno prostático específicoNanopartículas de ouroDetecção eletroquímicaLayer-by-Layer filmsanti-PSA antibodyProstate specific antigenGold nanoparticlesElectrochemical detectionCIENCIAS BIOLOGICAS::BIOQUIMICACIENCIAS DA SAUDE::MEDICINACIENCIAS DA SAUDE::MEDICINA::CLINICA MEDICA::CANCEROLOGIACIENCIAS EXATAS E DA TERRA::QUIMICA::FISICO-QUIMICA::ELETROQUIMICAFilmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouroNanostructured films for detection of the prostatic specific antigen using gold nanoparticlesinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline600600d379d7a0-e1eb-46b2-9712-8a73d0e062b4info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALDISSERTAÇÃO - Final 22-02-2019.pdfDISSERTAÇÃO - Final 22-02-2019.pdfapplication/pdf5740453https://repositorio.ufscar.br/bitstream/ufscar/11031/1/DISSERTAC%cc%a7A%cc%83O%20-%20Final%2022-02-2019.pdfba4dac100d097e5919069bc814c1788eMD51Carta comprovante.pdfCarta comprovante.pdfapplication/pdf3548567https://repositorio.ufscar.br/bitstream/ufscar/11031/2/Carta%20comprovante.pdf36d51a42131266d6067dbf69abf093adMD52LICENSElicense.txtlicense.txttext/plain; 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dc.title.por.fl_str_mv |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
dc.title.alternative.eng.fl_str_mv |
Nanostructured films for detection of the prostatic specific antigen using gold nanoparticles |
title |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
spellingShingle |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro Camilo, Douglas Eleutério Filmes camada por camada Anticorpo anti-PSA Antígeno prostático específico Nanopartículas de ouro Detecção eletroquímica Layer-by-Layer films anti-PSA antibody Prostate specific antigen Gold nanoparticles Electrochemical detection CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS DA SAUDE::MEDICINA CIENCIAS DA SAUDE::MEDICINA::CLINICA MEDICA::CANCEROLOGIA CIENCIAS EXATAS E DA TERRA::QUIMICA::FISICO-QUIMICA::ELETROQUIMICA |
title_short |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
title_full |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
title_fullStr |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
title_full_unstemmed |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
title_sort |
Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro |
author |
Camilo, Douglas Eleutério |
author_facet |
Camilo, Douglas Eleutério |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/1844596119185279 |
dc.contributor.author.fl_str_mv |
Camilo, Douglas Eleutério |
dc.contributor.advisor1.fl_str_mv |
Ferreira, Marystela |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/4714185516263478 |
dc.contributor.advisor-co1.fl_str_mv |
Massumi Miyazaki, Celina |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/2932962321837120 |
dc.contributor.authorID.fl_str_mv |
50fc25b9-3a47-4a4a-9d23-b936eca9185c |
contributor_str_mv |
Ferreira, Marystela Massumi Miyazaki, Celina |
dc.subject.por.fl_str_mv |
Filmes camada por camada Anticorpo anti-PSA Antígeno prostático específico Nanopartículas de ouro Detecção eletroquímica Layer-by-Layer films anti-PSA antibody Prostate specific antigen |
topic |
Filmes camada por camada Anticorpo anti-PSA Antígeno prostático específico Nanopartículas de ouro Detecção eletroquímica Layer-by-Layer films anti-PSA antibody Prostate specific antigen Gold nanoparticles Electrochemical detection CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS DA SAUDE::MEDICINA CIENCIAS DA SAUDE::MEDICINA::CLINICA MEDICA::CANCEROLOGIA CIENCIAS EXATAS E DA TERRA::QUIMICA::FISICO-QUIMICA::ELETROQUIMICA |
dc.subject.eng.fl_str_mv |
Gold nanoparticles Electrochemical detection |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS::BIOQUIMICA CIENCIAS DA SAUDE::MEDICINA CIENCIAS DA SAUDE::MEDICINA::CLINICA MEDICA::CANCEROLOGIA CIENCIAS EXATAS E DA TERRA::QUIMICA::FISICO-QUIMICA::ELETROQUIMICA |
description |
In the present work, nanostructured films were prepared through the self-assembly techniques for the detection of prostatic specific antigen (PSA), a glycoprotein produced in abnormal levels in case of prostate cancer. Two self-assembly approaches were used: the self-assembled monolayer (SAM) technique for the substrate functionalization and the layer-by-layer (LbL) technique for the building of the sensing unit. The specific antibody to PSA (anti-PSA) was immobilized on the gold nanoparticles surface (AuNP-antiPSA conjugate) for later assembly by the LbL technique. The AuNPs-anti-PSA conjugate was characterized by ultraviolet-visible spectroscopy indicating absorption bands at 520 and 260 nm related to the AuNP plasmons absorption and anti-PSA tryptophan absorption, respectively. The fluorescence spectrum presented a band at 367 nm attributed to the emission of tryptophan fluorescence present in the anti-PSA. The morphology analysis of AuNPs by Transmission Electron Microscopy (MET) provided an estimated average diameter of 3.5 ± 0.9 nm. The films were fabricated on gold (Au) covered glass slides previously cleaned and functionalized with 11-mercaptoundecanoic acid (11-MUA). The LbL films were prepared by the sequential adsorption of nanostructured layers of poly(ethyleneimine) (PEI), poly(vinylsulfonic acid, sodium salt) (PVS) and PEI, on the Au functionalized surface. Cyclic voltammetry (VC) confirmed the adsorption of the layers by the reduction of the current peak of Fe(II)/Fe(III) redox probe. Subsequently, bovine serum albumin (BSA) was used to block the non-specific binding sites. Detection was performed with different concentrations of PSA (0.1, 0.5, 1,0, 4,0, 8,0, 10 and 20 ng mL-1) using Electrochemical Impedance Spectroscopy (EIS). The architecture Au/11-MUA/PEI-PVS-PEI/AuNPs-anti-PSA/BSA/PSA was also characterized by Polarization-Modulation Infrared Reflection-Absorption Spectroscopy (PM-IRRAS) to investigate the interaction between the materials, as well as the interaction between antigen and antibody. The high selectivity of the immunosensor was tested by the exposure of the sensor to a non-specific immunoglobulin G (IgG). The limit of detection (LD) and the range of detection obtained by EIS were 0.17 ng mL-1 and 0.1-20 ng mL-1, respectively. The experiments demonstrated that the LbL technique and the use of AuNPs were suitable for the construction of an immunosensor of low cost, rapid development and high sensitivity. Thus, the fabricated immunoassay exhibits high potential for use in the diagnosis of the early stages of prostate cancer. |
publishDate |
2019 |
dc.date.accessioned.fl_str_mv |
2019-02-26T18:16:55Z |
dc.date.available.fl_str_mv |
2019-02-26T18:16:55Z |
dc.date.issued.fl_str_mv |
2019-02-15 |
dc.type.status.fl_str_mv |
info:eu-repo/semantics/publishedVersion |
dc.type.driver.fl_str_mv |
info:eu-repo/semantics/masterThesis |
format |
masterThesis |
status_str |
publishedVersion |
dc.identifier.citation.fl_str_mv |
CAMILO, Douglas Eleutério. Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro. 2019. Dissertação (Mestrado em Ciência dos Materiais) – Universidade Federal de São Carlos, Sorocaba, 2019. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11031. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/11031 |
identifier_str_mv |
CAMILO, Douglas Eleutério. Filmes nanoestruturados para detecção do antígeno prostático específico utilizando nanopartículas de ouro. 2019. Dissertação (Mestrado em Ciência dos Materiais) – Universidade Federal de São Carlos, Sorocaba, 2019. Disponível em: https://repositorio.ufscar.br/handle/ufscar/11031. |
url |
https://repositorio.ufscar.br/handle/ufscar/11031 |
dc.language.iso.fl_str_mv |
por |
language |
por |
dc.relation.confidence.fl_str_mv |
600 600 |
dc.relation.authority.fl_str_mv |
d379d7a0-e1eb-46b2-9712-8a73d0e062b4 |
dc.rights.driver.fl_str_mv |
info:eu-repo/semantics/openAccess |
eu_rights_str_mv |
openAccess |
dc.publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus Sorocaba |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Ciência dos Materiais - PPGCM-So |
dc.publisher.initials.fl_str_mv |
UFSCar |
publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus Sorocaba |
dc.source.none.fl_str_mv |
reponame:Repositório Institucional da UFSCAR instname:Universidade Federal de São Carlos (UFSCAR) instacron:UFSCAR |
instname_str |
Universidade Federal de São Carlos (UFSCAR) |
instacron_str |
UFSCAR |
institution |
UFSCAR |
reponame_str |
Repositório Institucional da UFSCAR |
collection |
Repositório Institucional da UFSCAR |
bitstream.url.fl_str_mv |
https://repositorio.ufscar.br/bitstream/ufscar/11031/1/DISSERTAC%cc%a7A%cc%83O%20-%20Final%2022-02-2019.pdf https://repositorio.ufscar.br/bitstream/ufscar/11031/2/Carta%20comprovante.pdf https://repositorio.ufscar.br/bitstream/ufscar/11031/3/license.txt https://repositorio.ufscar.br/bitstream/ufscar/11031/4/DISSERTAC%cc%a7A%cc%83O%20-%20Final%2022-02-2019.pdf.txt https://repositorio.ufscar.br/bitstream/ufscar/11031/5/Carta%20comprovante.pdf.txt https://repositorio.ufscar.br/bitstream/ufscar/11031/6/DISSERTAC%cc%a7A%cc%83O%20-%20Final%2022-02-2019.pdf.jpg https://repositorio.ufscar.br/bitstream/ufscar/11031/7/Carta%20comprovante.pdf.jpg |
bitstream.checksum.fl_str_mv |
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bitstream.checksumAlgorithm.fl_str_mv |
MD5 MD5 MD5 MD5 MD5 MD5 MD5 |
repository.name.fl_str_mv |
Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR) |
repository.mail.fl_str_mv |
|
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1802136352872988672 |