Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system

Autores
Arevalo, Fernando Javier; Messina, Germán Alejandro; Molina, Patricia Gabriela; Zon, María Alicia; Raba, Julio; Fernández, Héctor
Año de publicación
2010
Idioma
inglés
Tipo de recurso
artículo
Estado
versión publicada
Descripción
Progesterone (P4) is a steroidal hormone with a vital role in the maintenance of human and animal health. This paper describes the development of an immunosensor coupled to glassy carbon (GC) electrode and integrated to a microfluidic system to quantify P4 from bovine serum samples in a fast and sensitive way. The serum samples spiked with a given P4 concentration and a given P4 concentration bound to horseradish peroxide (HPR) were simultaneously added and, therefore, they competed immunologically with sheep monoclonal anti-P4 antibodies that were immobilized at a rotating disk. HRP in the presence of hydrogen peroxide (H2O2) catalyzes the chatecol (H2Q) oxidation to benzoquinone (Q). Its reverse electrochemical reduction to H2Q can be detected at a GC electrode surface at -0.15 V by chronoamperometric measurements. These current responses are proportional to the enzyme activity and inversely proportional to the P4 amount present in bovine serum samples. This P4 immunosensor showed a linear working range from 0.5 to 12.5 ng mL-1. The detection (DL) and quantification (QL) limits were 0.2 and 0.5 ng mL-1, respectively. The electrochemical immunosensor had a higher sensitivity than the ELISA method using conventional spectrophotometric detections. However, both methods allowed us to obtain similar detection limits. The immunosensor allowed us to make up to 100 determinations on different samples without any previous pre-treatment. This behavior proved to be suitable to detect P4 in routine veterinary, clinical, biological, physiological, and analytical assays.
Fil: Fernando Javier Arévalo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina
Fil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina; Argentina
Fil: Molina, Patricia Gabriela. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; Argentina
Fil: Zon, María Alicia. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; Argentina
Fil: Raba, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina; Argentina
Fil: Héctor Fernández. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; Argentina
Materia
AMPEROMETRIC IMMUNOSENSOR
ENZYME IMMUNOASSAYS
FLOW INJECTION ANALYSIS
GLASSY CARBON ELECTRODE
HORSERADISH PEROXIDASE
PROGESTERONE
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:11336/92850
Acceder
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oai_identifier_str oai:ri.conicet.gov.ar:11336/92850
network_acronym_str CONICETDig
repository_id_str 3498
network_name_str CONICET Digital (CONICET)
spelling Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor systemArevalo, Fernando JavierMessina, Germán AlejandroMolina, Patricia GabrielaZon, María AliciaRaba, JulioFernández, HéctorAMPEROMETRIC IMMUNOSENSORENZYME IMMUNOASSAYSFLOW INJECTION ANALYSISGLASSY CARBON ELECTRODEHORSERADISH PEROXIDASEPROGESTERONEhttps://purl.org/becyt/ford/1.4https://purl.org/becyt/ford/1Progesterone (P4) is a steroidal hormone with a vital role in the maintenance of human and animal health. This paper describes the development of an immunosensor coupled to glassy carbon (GC) electrode and integrated to a microfluidic system to quantify P4 from bovine serum samples in a fast and sensitive way. The serum samples spiked with a given P4 concentration and a given P4 concentration bound to horseradish peroxide (HPR) were simultaneously added and, therefore, they competed immunologically with sheep monoclonal anti-P4 antibodies that were immobilized at a rotating disk. HRP in the presence of hydrogen peroxide (H2O2) catalyzes the chatecol (H2Q) oxidation to benzoquinone (Q). Its reverse electrochemical reduction to H2Q can be detected at a GC electrode surface at -0.15 V by chronoamperometric measurements. These current responses are proportional to the enzyme activity and inversely proportional to the P4 amount present in bovine serum samples. This P4 immunosensor showed a linear working range from 0.5 to 12.5 ng mL-1. The detection (DL) and quantification (QL) limits were 0.2 and 0.5 ng mL-1, respectively. The electrochemical immunosensor had a higher sensitivity than the ELISA method using conventional spectrophotometric detections. However, both methods allowed us to obtain similar detection limits. The immunosensor allowed us to make up to 100 determinations on different samples without any previous pre-treatment. This behavior proved to be suitable to detect P4 in routine veterinary, clinical, biological, physiological, and analytical assays.Fil: Fernando Javier Arévalo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; ArgentinaFil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina; ArgentinaFil: Molina, Patricia Gabriela. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; ArgentinaFil: Zon, María Alicia. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; ArgentinaFil: Raba, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina; ArgentinaFil: Héctor Fernández. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; ArgentinaElsevier Science2010-03info:eu-repo/semantics/articleinfo:eu-repo/semantics/publishedVersionhttp://purl.org/coar/resource_type/c_6501info:ar-repo/semantics/articuloapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfapplication/pdfhttp://hdl.handle.net/11336/92850Arevalo, Fernando Javier; Messina, Germán Alejandro; Molina, Patricia Gabriela; Zon, María Alicia; Raba, Julio; et al.; Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system; Elsevier Science; Talanta; 80; 5; 3-2010; 1986-19920039-9140CONICET DigitalCONICETenginfo:eu-repo/semantics/altIdentifier/doi/10.1016/j.talanta.2009.10.059info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0039914009008480info:eu-repo/semantics/openAccesshttps://creativecommons.org/licenses/by-nc-sa/2.5/ar/reponame:CONICET Digital (CONICET)instname:Consejo Nacional de Investigaciones Científicas y Técnicas2025-09-03T09:43:26Zoai:ri.conicet.gov.ar:11336/92850instacron: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:34982025-09-03 09:43:26.751CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicasfalse
dc.title.none.fl_str_mv Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
title Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
spellingShingle Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
Arevalo, Fernando Javier
AMPEROMETRIC IMMUNOSENSOR
ENZYME IMMUNOASSAYS
FLOW INJECTION ANALYSIS
GLASSY CARBON ELECTRODE
HORSERADISH PEROXIDASE
PROGESTERONE
title_short Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
title_full Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
title_fullStr Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
title_full_unstemmed Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
title_sort Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system
dc.creator.none.fl_str_mv Arevalo, Fernando Javier
Messina, Germán Alejandro
Molina, Patricia Gabriela
Zon, María Alicia
Raba, Julio
Fernández, Héctor
author Arevalo, Fernando Javier
author_facet Arevalo, Fernando Javier
Messina, Germán Alejandro
Molina, Patricia Gabriela
Zon, María Alicia
Raba, Julio
Fernández, Héctor
author_role author
author2 Messina, Germán Alejandro
Molina, Patricia Gabriela
Zon, María Alicia
Raba, Julio
Fernández, Héctor
author2_role author
author
author
author
author
dc.subject.none.fl_str_mv AMPEROMETRIC IMMUNOSENSOR
ENZYME IMMUNOASSAYS
FLOW INJECTION ANALYSIS
GLASSY CARBON ELECTRODE
HORSERADISH PEROXIDASE
PROGESTERONE
topic AMPEROMETRIC IMMUNOSENSOR
ENZYME IMMUNOASSAYS
FLOW INJECTION ANALYSIS
GLASSY CARBON ELECTRODE
HORSERADISH PEROXIDASE
PROGESTERONE
purl_subject.fl_str_mv https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
dc.description.none.fl_txt_mv Progesterone (P4) is a steroidal hormone with a vital role in the maintenance of human and animal health. This paper describes the development of an immunosensor coupled to glassy carbon (GC) electrode and integrated to a microfluidic system to quantify P4 from bovine serum samples in a fast and sensitive way. The serum samples spiked with a given P4 concentration and a given P4 concentration bound to horseradish peroxide (HPR) were simultaneously added and, therefore, they competed immunologically with sheep monoclonal anti-P4 antibodies that were immobilized at a rotating disk. HRP in the presence of hydrogen peroxide (H2O2) catalyzes the chatecol (H2Q) oxidation to benzoquinone (Q). Its reverse electrochemical reduction to H2Q can be detected at a GC electrode surface at -0.15 V by chronoamperometric measurements. These current responses are proportional to the enzyme activity and inversely proportional to the P4 amount present in bovine serum samples. This P4 immunosensor showed a linear working range from 0.5 to 12.5 ng mL-1. The detection (DL) and quantification (QL) limits were 0.2 and 0.5 ng mL-1, respectively. The electrochemical immunosensor had a higher sensitivity than the ELISA method using conventional spectrophotometric detections. However, both methods allowed us to obtain similar detection limits. The immunosensor allowed us to make up to 100 determinations on different samples without any previous pre-treatment. This behavior proved to be suitable to detect P4 in routine veterinary, clinical, biological, physiological, and analytical assays.
Fil: Fernando Javier Arévalo. Universidad Nacional de Río Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales; Argentina
Fil: Messina, Germán Alejandro. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina; Argentina
Fil: Molina, Patricia Gabriela. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; Argentina
Fil: Zon, María Alicia. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; Argentina
Fil: Raba, Julio. Consejo Nacional de Investigaciones Científicas y Técnicas. Centro Científico Tecnológico Conicet - San Luis. Instituto de Química de San Luis. Universidad Nacional de San Luis. Facultad de Química, Bioquímica y Farmacia. Instituto de Química de San Luis; Argentina; Argentina
Fil: Héctor Fernández. Universidad Nacional de Rio Cuarto. Facultad de Ciencias Exactas Fisicoquímicas y Naturales. Departamento de Química. Area Electroquímica; Argentina
description Progesterone (P4) is a steroidal hormone with a vital role in the maintenance of human and animal health. This paper describes the development of an immunosensor coupled to glassy carbon (GC) electrode and integrated to a microfluidic system to quantify P4 from bovine serum samples in a fast and sensitive way. The serum samples spiked with a given P4 concentration and a given P4 concentration bound to horseradish peroxide (HPR) were simultaneously added and, therefore, they competed immunologically with sheep monoclonal anti-P4 antibodies that were immobilized at a rotating disk. HRP in the presence of hydrogen peroxide (H2O2) catalyzes the chatecol (H2Q) oxidation to benzoquinone (Q). Its reverse electrochemical reduction to H2Q can be detected at a GC electrode surface at -0.15 V by chronoamperometric measurements. These current responses are proportional to the enzyme activity and inversely proportional to the P4 amount present in bovine serum samples. This P4 immunosensor showed a linear working range from 0.5 to 12.5 ng mL-1. The detection (DL) and quantification (QL) limits were 0.2 and 0.5 ng mL-1, respectively. The electrochemical immunosensor had a higher sensitivity than the ELISA method using conventional spectrophotometric detections. However, both methods allowed us to obtain similar detection limits. The immunosensor allowed us to make up to 100 determinations on different samples without any previous pre-treatment. This behavior proved to be suitable to detect P4 in routine veterinary, clinical, biological, physiological, and analytical assays.
publishDate 2010
dc.date.none.fl_str_mv 2010-03
dc.type.none.fl_str_mv info:eu-repo/semantics/article
info:eu-repo/semantics/publishedVersion
http://purl.org/coar/resource_type/c_6501
info:ar-repo/semantics/articulo
format article
status_str publishedVersion
dc.identifier.none.fl_str_mv http://hdl.handle.net/11336/92850
Arevalo, Fernando Javier; Messina, Germán Alejandro; Molina, Patricia Gabriela; Zon, María Alicia; Raba, Julio; et al.; Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system; Elsevier Science; Talanta; 80; 5; 3-2010; 1986-1992
0039-9140
CONICET Digital
CONICET
url http://hdl.handle.net/11336/92850
identifier_str_mv Arevalo, Fernando Javier; Messina, Germán Alejandro; Molina, Patricia Gabriela; Zon, María Alicia; Raba, Julio; et al.; Determination of progesterone (P4) from bovine serum samples using a microfluidic immunosensor system; Elsevier Science; Talanta; 80; 5; 3-2010; 1986-1992
0039-9140
CONICET Digital
CONICET
dc.language.none.fl_str_mv eng
language eng
dc.relation.none.fl_str_mv info:eu-repo/semantics/altIdentifier/doi/10.1016/j.talanta.2009.10.059
info:eu-repo/semantics/altIdentifier/url/https://www.sciencedirect.com/science/article/abs/pii/S0039914009008480
dc.rights.none.fl_str_mv info:eu-repo/semantics/openAccess
https://creativecommons.org/licenses/by-nc-sa/2.5/ar/
eu_rights_str_mv openAccess
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
application/pdf
application/pdf
application/pdf
dc.publisher.none.fl_str_mv Elsevier Science
publisher.none.fl_str_mv Elsevier Science
dc.source.none.fl_str_mv reponame:CONICET Digital (CONICET)
instname:Consejo Nacional de Investigaciones Científicas y Técnicas
reponame_str CONICET Digital (CONICET)
collection CONICET Digital (CONICET)
instname_str Consejo Nacional de Investigaciones Científicas y Técnicas
repository.name.fl_str_mv CONICET Digital (CONICET) - Consejo Nacional de Investigaciones Científicas y Técnicas
repository.mail.fl_str_mv dasensio@conicet.gov.ar; lcarlino@conicet.gov.ar
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score 13.13397

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