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

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 sen...

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Detalles Bibliográficos
Autores: Arevalo, Fernando Javier, Messina, Germán Alejandro, Molina, Patricia Gabriela, Zon, María Alicia, Raba, Julio, Fernández, Héctor
Tipo de recurso: artículo
Estado:Versión publicada
Fecha de publicación:2010
País:Argentina
Institución:Consejo Nacional de Investigaciones Científicas y Técnicas
Repositorio:CONICET Digital (CONICET)
Idioma:inglés
OAI Identifier:oai:ri.conicet.gov.ar:11336/92850
Acceso en línea:http://hdl.handle.net/11336/92850
Access Level:acceso abierto
Palabra clave:AMPEROMETRIC IMMUNOSENSOR
ENZYME IMMUNOASSAYS
FLOW INJECTION ANALYSIS
GLASSY CARBON ELECTRODE
HORSERADISH PEROXIDASE
PROGESTERONE
https://purl.org/becyt/ford/1.4
https://purl.org/becyt/ford/1
Descripción
Sumario: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.