Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta
Autor(a) principal: | |
---|---|
Data de Publicação: | 2016 |
Tipo de documento: | Dissertação |
Idioma: | por |
Título da fonte: | Repositório Institucional da UFSCAR |
Texto Completo: | https://repositorio.ufscar.br/handle/ufscar/8070 |
Resumo: | This study presents the development of a biosensor using a microcantilever stem and its detecting response through dynamic mode of atomic force microscopy (AFM). Responses of different methods for immobilization of alcohol dehydrogenase enzyme were investigated in order to accomplish a selective detection of ethanol, methanol and isopropanol, using as arrays: self-assembly monolayer (SAM) and others arrangements formed on Si and Si3N4 microcantilevers surface. In addition, we aimed to evaluate the influence of the biological element immobilization process in the analytical performance of the biosensor. The process to development of a biosensor began with the activation of its surface by means of thiols and silanes with different carbon length chains, however with the same amine terminal groups (NH2). The microcantilevers activation is only possible with the oxidation of its surface and to check this process, the "High-resolution Xray Photoelectron Spectroscopy (XPS)" technique was used. Activation of the surface was obtained using different methods for coating of microcantilevers made only with Si (100) and Si3N4. Subsequently to the oxidation process, the binding of the biomolecule to the surface activated was performed using the bifunctional agent glutaraldehyde. Microcantilevers with both sides coated by biomolecules were investigated and a tension effect of surface was observed at a 0,3 mL/L of the target analyte. By analyzing the performance of the biosensor for the determination of the target analyte, it was realized that APTES activation methodology for biosensors steam and self-assembled monolayers were the most suitable techniques for immobilizing the recognition biomolecule. With this preparation, the biosensor it showed less susceptible to humidity and the temperature variations, presenting a high quality factor, a faster response time, selectivity, sensitivity and durability. |
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Margarido, AlexandreHerrmann Júnior, Paulo Sérgio de Paulahttp://lattes.cnpq.br/3478230611350302Araujo-Moreira, Fernando M.http://lattes.cnpq.br/1809254923092721http://lattes.cnpq.br/124002594319363643fb1892-5797-4495-b48c-9fcb21d11ed22016-10-20T19:45:37Z2016-10-20T19:45:37Z2016-07-11MARGARIDO, Alexandre. Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta. 2016. Dissertação (Mestrado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8070.https://repositorio.ufscar.br/handle/ufscar/8070This study presents the development of a biosensor using a microcantilever stem and its detecting response through dynamic mode of atomic force microscopy (AFM). Responses of different methods for immobilization of alcohol dehydrogenase enzyme were investigated in order to accomplish a selective detection of ethanol, methanol and isopropanol, using as arrays: self-assembly monolayer (SAM) and others arrangements formed on Si and Si3N4 microcantilevers surface. In addition, we aimed to evaluate the influence of the biological element immobilization process in the analytical performance of the biosensor. The process to development of a biosensor began with the activation of its surface by means of thiols and silanes with different carbon length chains, however with the same amine terminal groups (NH2). The microcantilevers activation is only possible with the oxidation of its surface and to check this process, the "High-resolution Xray Photoelectron Spectroscopy (XPS)" technique was used. Activation of the surface was obtained using different methods for coating of microcantilevers made only with Si (100) and Si3N4. Subsequently to the oxidation process, the binding of the biomolecule to the surface activated was performed using the bifunctional agent glutaraldehyde. Microcantilevers with both sides coated by biomolecules were investigated and a tension effect of surface was observed at a 0,3 mL/L of the target analyte. By analyzing the performance of the biosensor for the determination of the target analyte, it was realized that APTES activation methodology for biosensors steam and self-assembled monolayers were the most suitable techniques for immobilizing the recognition biomolecule. With this preparation, the biosensor it showed less susceptible to humidity and the temperature variations, presenting a high quality factor, a faster response time, selectivity, sensitivity and durability.Este trabalho apresenta o desenvolvimento de um biossensor, utilizando a haste de microcantilever e a detecção de sua resposta por intermédio do modo dinâmico da microscopia de força atômica (AFM). Foram investigadas as respostas de diferentes métodos para imobilização da enzima Álcool Desidrogenase, a fim de realizar a detecção seletiva de etanol, metanol e isopropanol, empregando como matrizes monocamadas auto-organizadas (SAM – Self Assembed Monolayer) e outros arranjos formados sobre a superfície de microcantilevers de Si e Si3N4. Além disso, objetivou-se a avaliação da influência do processo de imobilização do elemento biológico no desempenho analítico do biossensor. O processo de desenvolvimento do biossensor iniciou-se com a ativação de sua superfície por meio de deposição de tióis e silanos com diferentes comprimentos de cadeias carbônicas, mas com os mesmos grupos terminais amina (NH2). A ativação de microcantilevers somente é possível com a oxidação de sua superfície e para verificação desse processo foi empregada a técnica “High-resolution Xray Photoelectron Spectroscopy (XPS)”. A ativação da superfície foi obtida utilizando metodologias diferentes para o recobrimento dos microcantilevers construídos somente com Si (100) e Si3N4. Posteriormente ao processo de oxidação, segue-se a ligação da biomolécula à superfície ativada pela utilização do agente bifuncional Glutaraldeído. Foram investigados microcantilevers em que ambos os lados foram revestidos com as biomoléculas e, foi constatado o efeito da tensão superficial a partir de 0,3 mL/L do analito alvo. Ao analisar o desempenho do biossensor para a determinação do analito alvo, verificou-se que nos biossensores a metodologia de ativação por vapor de (3-Aminopropyl)triethoxysilane (APTES) e com montagem em monocamadas auto-organizada foi a mais adequada para a imobilização da biomolécula de reconhecimento, com esta preparação o biossensor foi menos susceptível a umidade relativa e variações de temperatura apresentando elevado fator de qualidade, menor período de tempo de resposta, seletividade, durabilidade e sensibilidade.Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)porUniversidade Federal de São CarlosCâmpus São CarlosPrograma de Pós-Graduação em Biotecnologia - PPGBiotecUFSCarBiossensorMicroscópio de força atômicaÁlcool desidrogenaseTensão superficialMicrocantileverCIENCIAS BIOLOGICASDesenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curtainfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/masterThesisOnline600bc47636f-2f51-4b82-91f7-484701cb0784info:eu-repo/semantics/openAccessreponame:Repositório Institucional da UFSCARinstname:Universidade Federal de São Carlos (UFSCAR)instacron:UFSCARORIGINALTeseAM.pdfTeseAM.pdfapplication/pdf7680112https://repositorio.ufscar.br/bitstream/ufscar/8070/1/TeseAM.pdfc8ecf6ee83f4312fa3be9569904c7b2fMD51LICENSElicense.txtlicense.txttext/plain; charset=utf-81957https://repositorio.ufscar.br/bitstream/ufscar/8070/2/license.txtae0398b6f8b235e40ad82cba6c50031dMD52TEXTTeseAM.pdf.txtTeseAM.pdf.txtExtracted texttext/plain220619https://repositorio.ufscar.br/bitstream/ufscar/8070/3/TeseAM.pdf.txt1d9732b1346cb0e61723abdf68dfaec2MD53THUMBNAILTeseAM.pdf.jpgTeseAM.pdf.jpgIM Thumbnailimage/jpeg8923https://repositorio.ufscar.br/bitstream/ufscar/8070/4/TeseAM.pdf.jpgd6c741b5bae5997fb5f11ec4452ba0caMD54ufscar/80702023-09-18 18:31:09.465oai:repositorio.ufscar.br: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Repositório InstitucionalPUBhttps://repositorio.ufscar.br/oai/requestopendoar:43222023-09-18T18:31:09Repositório Institucional da UFSCAR - Universidade Federal de São Carlos (UFSCAR)false |
dc.title.por.fl_str_mv |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
title |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
spellingShingle |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta Margarido, Alexandre Biossensor Microscópio de força atômica Álcool desidrogenase Tensão superficial Microcantilever CIENCIAS BIOLOGICAS |
title_short |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
title_full |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
title_fullStr |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
title_full_unstemmed |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
title_sort |
Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta |
author |
Margarido, Alexandre |
author_facet |
Margarido, Alexandre |
author_role |
author |
dc.contributor.authorlattes.por.fl_str_mv |
http://lattes.cnpq.br/1240025943193636 |
dc.contributor.author.fl_str_mv |
Margarido, Alexandre |
dc.contributor.advisor1.fl_str_mv |
Herrmann Júnior, Paulo Sérgio de Paula |
dc.contributor.advisor1Lattes.fl_str_mv |
http://lattes.cnpq.br/3478230611350302 |
dc.contributor.advisor-co1.fl_str_mv |
Araujo-Moreira, Fernando M. |
dc.contributor.advisor-co1Lattes.fl_str_mv |
http://lattes.cnpq.br/1809254923092721 |
dc.contributor.authorID.fl_str_mv |
43fb1892-5797-4495-b48c-9fcb21d11ed2 |
contributor_str_mv |
Herrmann Júnior, Paulo Sérgio de Paula Araujo-Moreira, Fernando M. |
dc.subject.por.fl_str_mv |
Biossensor Microscópio de força atômica Álcool desidrogenase Tensão superficial Microcantilever |
topic |
Biossensor Microscópio de força atômica Álcool desidrogenase Tensão superficial Microcantilever CIENCIAS BIOLOGICAS |
dc.subject.cnpq.fl_str_mv |
CIENCIAS BIOLOGICAS |
description |
This study presents the development of a biosensor using a microcantilever stem and its detecting response through dynamic mode of atomic force microscopy (AFM). Responses of different methods for immobilization of alcohol dehydrogenase enzyme were investigated in order to accomplish a selective detection of ethanol, methanol and isopropanol, using as arrays: self-assembly monolayer (SAM) and others arrangements formed on Si and Si3N4 microcantilevers surface. In addition, we aimed to evaluate the influence of the biological element immobilization process in the analytical performance of the biosensor. The process to development of a biosensor began with the activation of its surface by means of thiols and silanes with different carbon length chains, however with the same amine terminal groups (NH2). The microcantilevers activation is only possible with the oxidation of its surface and to check this process, the "High-resolution Xray Photoelectron Spectroscopy (XPS)" technique was used. Activation of the surface was obtained using different methods for coating of microcantilevers made only with Si (100) and Si3N4. Subsequently to the oxidation process, the binding of the biomolecule to the surface activated was performed using the bifunctional agent glutaraldehyde. Microcantilevers with both sides coated by biomolecules were investigated and a tension effect of surface was observed at a 0,3 mL/L of the target analyte. By analyzing the performance of the biosensor for the determination of the target analyte, it was realized that APTES activation methodology for biosensors steam and self-assembled monolayers were the most suitable techniques for immobilizing the recognition biomolecule. With this preparation, the biosensor it showed less susceptible to humidity and the temperature variations, presenting a high quality factor, a faster response time, selectivity, sensitivity and durability. |
publishDate |
2016 |
dc.date.accessioned.fl_str_mv |
2016-10-20T19:45:37Z |
dc.date.available.fl_str_mv |
2016-10-20T19:45:37Z |
dc.date.issued.fl_str_mv |
2016-07-11 |
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 |
MARGARIDO, Alexandre. Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta. 2016. Dissertação (Mestrado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8070. |
dc.identifier.uri.fl_str_mv |
https://repositorio.ufscar.br/handle/ufscar/8070 |
identifier_str_mv |
MARGARIDO, Alexandre. Desenvolvimento de biossensor baseado em microcantilever funcionalizado com biomoléculas para a detecção de alcoois de cadeia curta. 2016. Dissertação (Mestrado em Biotecnologia) – Universidade Federal de São Carlos, São Carlos, 2016. Disponível em: https://repositorio.ufscar.br/handle/ufscar/8070. |
url |
https://repositorio.ufscar.br/handle/ufscar/8070 |
dc.language.iso.fl_str_mv |
por |
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por |
dc.relation.confidence.fl_str_mv |
600 |
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bc47636f-2f51-4b82-91f7-484701cb0784 |
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 São Carlos |
dc.publisher.program.fl_str_mv |
Programa de Pós-Graduação em Biotecnologia - PPGBiotec |
dc.publisher.initials.fl_str_mv |
UFSCar |
publisher.none.fl_str_mv |
Universidade Federal de São Carlos Câmpus São Carlos |
dc.source.none.fl_str_mv |
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