Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor

Autores
Rodriguez, Carolina Mercedes; Brito Hoyos, Diana Marcela; Perillo, Maria Angelica; Clop, Eduardo Matias
Año de publicación
2022
Idioma
inglés
Tipo de recurso
documento de conferencia
Estado
versión publicada
Descripción
Many biological phenomena can be characterized through the use of biosensors. The use of enzymes as a biological component allows to achieve specificity and speed in measurements. In our laboratory we have been working on the design of a biosensor based on the activity of bovine erythrocyte acetylcholinesterase BEA, testing the effect of different compounds on the catalytic activity of the enzyme. To do this, Langmuir films (LF) build up from bovine erythrocyte membrane (BEM) are transferred to an alkylated glass substrate through the Langmuir Schaefer method (LS films). To improve the reproducibility of results and enhance the biosensor efficiency, different techniques were assayed. for LF preparation. Membrane spreading was done applying drop by drop deposition over the air-water interface or drop slippering up to the interface along a glass rod. Moreover, the film transference from the air-water interface to the glass substrate was done with or without the elimination, by aspiration, of the monolayer not adhered to the glass. The catalytic activity of BEA and the protein surface density were determined at 96 discrete points covering the entire surface of the glass substrate. These data were plotted as a 3D mesh and submitted to a statistical analysis using the Moran’s index to evaluate the film homogeneity. In some cases, the LS film was also prepared with a trace amount of the dye DiI-C18 and the fluorescence intensity was scanned along the whole plate using an Odissey scanner. At the macroscopic level, fluorescence images exhibited a homogeneous film. The biochemical data exhibited a varied degree of heterogeneity distribution in the 3D mesh. However, statistical analysis using Moran's index showed that the values consisted of almost randomly distributed spatial data, with slight variations depending on the technique applied to the LF and LS preparation. Moreover, the least coefficient of variations were obtained with rod spreading without interface aspiration. It can be useful to take into account possible disturbances in the monolayer organization due to the dynamics followed by the catenoidal shaped capillary bridge formed between the solid substrate and the liquid interface during the elevation of the LS substrate. To get deeply into this problem, modifications in the substrate design will be assayed.
Fil: Rodriguez, Carolina Mercedes. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Brito Hoyos, Diana Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Clop, Eduardo Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
L Reunión Anual Sociedad Argentina de Biofísica
Rosario
Argentina
Sociedad Argentina de Biofísica
Materia
Biosensor
acetilcolinestersa de eritrocito bovino
Biofisica de membranas
Enzyme kinetics
Nivel de accesibilidad
acceso abierto
Condiciones de uso
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
Repositorio
CONICET Digital (CONICET)
Institución
Consejo Nacional de Investigaciones Científicas y Técnicas
OAI Identificador
oai:ri.conicet.gov.ar:11336/233769
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/233769
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repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensorRodriguez, Carolina MercedesBrito Hoyos, Diana MarcelaPerillo, Maria AngelicaClop, Eduardo MatiasBiosensoracetilcolinestersa de eritrocito bovinoBiofisica de membranasEnzyme kineticshttps://purl.org/becyt/ford/1.6https://purl.org/becyt/ford/1Many biological phenomena can be characterized through the use of biosensors. The use of enzymes as a biological component allows to achieve specificity and speed in measurements. In our laboratory we have been working on the design of a biosensor based on the activity of bovine erythrocyte acetylcholinesterase BEA, testing the effect of different compounds on the catalytic activity of the enzyme. To do this, Langmuir films (LF) build up from bovine erythrocyte membrane (BEM) are transferred to an alkylated glass substrate through the Langmuir Schaefer method (LS films). To improve the reproducibility of results and enhance the biosensor efficiency, different techniques were assayed. for LF preparation. Membrane spreading was done applying drop by drop deposition over the air-water interface or drop slippering up to the interface along a glass rod. Moreover, the film transference from the air-water interface to the glass substrate was done with or without the elimination, by aspiration, of the monolayer not adhered to the glass. The catalytic activity of BEA and the protein surface density were determined at 96 discrete points covering the entire surface of the glass substrate. These data were plotted as a 3D mesh and submitted to a statistical analysis using the Moran’s index to evaluate the film homogeneity. In some cases, the LS film was also prepared with a trace amount of the dye DiI-C18 and the fluorescence intensity was scanned along the whole plate using an Odissey scanner. At the macroscopic level, fluorescence images exhibited a homogeneous film. The biochemical data exhibited a varied degree of heterogeneity distribution in the 3D mesh. However, statistical analysis using Moran's index showed that the values consisted of almost randomly distributed spatial data, with slight variations depending on the technique applied to the LF and LS preparation. Moreover, the least coefficient of variations were obtained with rod spreading without interface aspiration. It can be useful to take into account possible disturbances in the monolayer organization due to the dynamics followed by the catenoidal shaped capillary bridge formed between the solid substrate and the liquid interface during the elevation of the LS substrate. To get deeply into this problem, modifications in the substrate design will be assayed.Fil: Rodriguez, Carolina Mercedes. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Brito Hoyos, Diana Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; ArgentinaFil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaFil: Clop, Eduardo Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; ArgentinaL Reunión Anual Sociedad Argentina de BiofísicaRosarioArgentinaSociedad Argentina de BiofísicaSociedad Argentina de BiofísicaAmbroggio, Ernesto EstebanCelej, Maria SoledadAcierno, Juan Pablo2022info:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/conferenceObjectReuniónBookhttp://purl.org/coar/resource_type/c_5794info:ar-repo/semantics/documentoDeConferenciaapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/233769Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor; L Reunión Anual Sociedad Argentina de Biofísica; Rosario; Argentina; 2022; 205-205978-987-48938-0-2CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/url/https://biofisica.org.ar/reuniones-cientificas/reunionsab-previas/Nacionalinfo:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-12-23T14:46:41Zoai:ri.conicet.gov.ar:11336/233769instacron:CONICETInstitucionalhttp://ri.conicet.gov.ar/Organismo científico-tecnológicoNo correspondehttp://ri.conicet.gov.ar/oai/requestdasensio@conicet.gov.ar; lcarlino@conicet.gov.arArgentinaNo correspondeNo correspondeNo correspondeopendoar:34982025-12-23 14:46:41.514CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
title Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
spellingShingle Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
Rodriguez, Carolina Mercedes
Biosensor
acetilcolinestersa de eritrocito bovino
Biofisica de membranas
Enzyme kinetics
title_short Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
title_full Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
title_fullStr Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
title_full_unstemmed Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
title_sort Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor
dc.creator.none.fl_str_mv Rodriguez, Carolina Mercedes
Brito Hoyos, Diana Marcela
Perillo, Maria Angelica
Clop, Eduardo Matias
author Rodriguez, Carolina Mercedes
author_facet Rodriguez, Carolina Mercedes
Brito Hoyos, Diana Marcela
Perillo, Maria Angelica
Clop, Eduardo Matias
author_role author
author2 Brito Hoyos, Diana Marcela
Perillo, Maria Angelica
Clop, Eduardo Matias
author2_role author
author
author
dc.contributor.none.fl_str_mv Ambroggio, Ernesto Esteban
Celej, Maria Soledad
Acierno, Juan Pablo
dc.subject.none.fl_str_mv Biosensor
acetilcolinestersa de eritrocito bovino
Biofisica de membranas
Enzyme kinetics
topic Biosensor
acetilcolinestersa de eritrocito bovino
Biofisica de membranas
Enzyme kinetics
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.6
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Many biological phenomena can be characterized through the use of biosensors. The use of enzymes as a biological component allows to achieve specificity and speed in measurements. In our laboratory we have been working on the design of a biosensor based on the activity of bovine erythrocyte acetylcholinesterase BEA, testing the effect of different compounds on the catalytic activity of the enzyme. To do this, Langmuir films (LF) build up from bovine erythrocyte membrane (BEM) are transferred to an alkylated glass substrate through the Langmuir Schaefer method (LS films). To improve the reproducibility of results and enhance the biosensor efficiency, different techniques were assayed. for LF preparation. Membrane spreading was done applying drop by drop deposition over the air-water interface or drop slippering up to the interface along a glass rod. Moreover, the film transference from the air-water interface to the glass substrate was done with or without the elimination, by aspiration, of the monolayer not adhered to the glass. The catalytic activity of BEA and the protein surface density were determined at 96 discrete points covering the entire surface of the glass substrate. These data were plotted as a 3D mesh and submitted to a statistical analysis using the Moran’s index to evaluate the film homogeneity. In some cases, the LS film was also prepared with a trace amount of the dye DiI-C18 and the fluorescence intensity was scanned along the whole plate using an Odissey scanner. At the macroscopic level, fluorescence images exhibited a homogeneous film. The biochemical data exhibited a varied degree of heterogeneity distribution in the 3D mesh. However, statistical analysis using Moran's index showed that the values consisted of almost randomly distributed spatial data, with slight variations depending on the technique applied to the LF and LS preparation. Moreover, the least coefficient of variations were obtained with rod spreading without interface aspiration. It can be useful to take into account possible disturbances in the monolayer organization due to the dynamics followed by the catenoidal shaped capillary bridge formed between the solid substrate and the liquid interface during the elevation of the LS substrate. To get deeply into this problem, modifications in the substrate design will be assayed.
Fil: Rodriguez, Carolina Mercedes. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Brito Hoyos, Diana Marcela. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina
Fil: Perillo, Maria Angelica. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
Fil: Clop, Eduardo Matias. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - Córdoba. Instituto de Investigaciones Biológicas y Tecnológicas. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Instituto de Investigaciones Biológicas y Tecnológicas; Argentina. Universidad Nacional de Córdoba. Facultad de Ciencias Exactas, Físicas y Naturales. Departamento de Química. Cátedra de Química Biológica; Argentina
L Reunión Anual Sociedad Argentina de Biofísica
Rosario
Argentina
Sociedad Argentina de Biofísica
description Many biological phenomena can be characterized through the use of biosensors. The use of enzymes as a biological component allows to achieve specificity and speed in measurements. In our laboratory we have been working on the design of a biosensor based on the activity of bovine erythrocyte acetylcholinesterase BEA, testing the effect of different compounds on the catalytic activity of the enzyme. To do this, Langmuir films (LF) build up from bovine erythrocyte membrane (BEM) are transferred to an alkylated glass substrate through the Langmuir Schaefer method (LS films). To improve the reproducibility of results and enhance the biosensor efficiency, different techniques were assayed. for LF preparation. Membrane spreading was done applying drop by drop deposition over the air-water interface or drop slippering up to the interface along a glass rod. Moreover, the film transference from the air-water interface to the glass substrate was done with or without the elimination, by aspiration, of the monolayer not adhered to the glass. The catalytic activity of BEA and the protein surface density were determined at 96 discrete points covering the entire surface of the glass substrate. These data were plotted as a 3D mesh and submitted to a statistical analysis using the Moran’s index to evaluate the film homogeneity. In some cases, the LS film was also prepared with a trace amount of the dye DiI-C18 and the fluorescence intensity was scanned along the whole plate using an Odissey scanner. At the macroscopic level, fluorescence images exhibited a homogeneous film. The biochemical data exhibited a varied degree of heterogeneity distribution in the 3D mesh. However, statistical analysis using Moran's index showed that the values consisted of almost randomly distributed spatial data, with slight variations depending on the technique applied to the LF and LS preparation. Moreover, the least coefficient of variations were obtained with rod spreading without interface aspiration. It can be useful to take into account possible disturbances in the monolayer organization due to the dynamics followed by the catenoidal shaped capillary bridge formed between the solid substrate and the liquid interface during the elevation of the LS substrate. To get deeply into this problem, modifications in the substrate design will be assayed.
publishDate 2022
dc.date.none.fl_str_mv 2022
dc.type.none.fl_str_mv info:eu-repo/semantics/publishedVersion
info:eu-repo/semantics/conferenceObject
Reunión
Book
http://purl.org/coar/resource_type/c_5794
info:ar-repo/semantics/documentoDeConferencia
status_str publishedVersion
format conferenceObject
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/233769
Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor; L Reunión Anual Sociedad Argentina de Biofísica; Rosario; Argentina; 2022; 205-205
978-987-48938-0-2
CONICET Digital
CONICET
url http://hdl.handle.net/11336/233769
identifier_str_mv Design and development of a bovine erythrocyte acetylcholinesterase (BEA) activity-based biosensor; L Reunión Anual Sociedad Argentina de Biofísica; Rosario; Argentina; 2022; 205-205
978-987-48938-0-2
CONICET Digital
CONICET
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language eng
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https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
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rights_invalid_str_mv https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
dc.format.none.fl_str_mv application/pdf
application/pdf
application/pdf
dc.coverage.none.fl_str_mv Nacional
dc.publisher.none.fl_str_mv Sociedad Argentina de Biofísica
publisher.none.fl_str_mv Sociedad Argentina de Biofísica
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