Lactate biosensing based on covalent immobilization of lactate oxidase onto chevron-like graphene nanoribbons via diazotization-coupling reaction

We have designed and prepared an electrochemical biosensor for lactate determination. Through a diazotation process, the enzyme lactate oxidase (LOx) is anchored onto chevron-like graphene nanoribbons (GNR), previously synthesized by a solution-based chemical route, and used as modifiers of glassy c...

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Detalles Bibliográficos
Autores: Sainz Vaque, Raquel, Pozo Vázquez, María del, Vazquez, Luís, Vilas Varela, Manuel, Castro Esteban, Jesús, Blanco Gil, Elías, Petit Domínguez, María Dolores, Quintana Mani, María del Carmen, Casero Junquera, María Elena
Tipo de recurso: artículo
Fecha de publicación:2022
País:España
Institución:Universidad Autónoma de Madrid
Repositorio:Biblos-e Archivo. Repositorio Institucional de la UAM
Idioma:inglés
OAI Identifier:oai:repositorio.uam.es:10486/703175
Acceso en línea:http://hdl.handle.net/10486/703175
https://dx.doi.org/10.1016/j.aca.2022.339851
Access Level:acceso abierto
Palabra clave:Biosensor
Chevron-like graphene nanoribbons
Lactate oxidase
Diazonium chemistry
Electrochemical detection
Lactate
Química
Descripción
Sumario:We have designed and prepared an electrochemical biosensor for lactate determination. Through a diazotation process, the enzyme lactate oxidase (LOx) is anchored onto chevron-like graphene nanoribbons (GNR), previously synthesized by a solution-based chemical route, and used as modifiers of glassy carbon electrodes. In a first step, we have performed the grafting of a 4-carboxyphenyl film, by electrochemical reduction of the corresponding 4-carboxyphenyl diazonium salt, on the GNR-modified electrode surface. In this way, the carboxylic groups are exposed to the solution, enabling the covalent immobilization of the enzyme through the formation of an amide bond between these carboxylic groups and the amine groups of the enzyme. The biosensor design was optimized through the morphological and electrochemical characterization of each construction step by atomic force microscopy, scanning electron microscopy, cyclic voltammetry and electrochemical impedance spectroscopy.The cyclic voltammetric response of the biosensor in a solution of hydroxymethylferrocene in presence of Llactate evidenced a clear electrocatalytic effect powered by the specific design of the biosensing platform with LOx covalently attached to the GNR layer. From the calibration procedures employed for L-lactate determination, a linear concentration range of 3.4 ⋅ 10− 5 – 2.8 ⋅ 10− 4 M and a detection limit of 11 μM were obtained, with relative errors and relative standard deviations less than 6.0% and 8.4%, respectively. The applicability of the biosensor was tested by determining lactate in apple juices, leading to results that are in good agreement with those obtained with a well-established enzymatic spectrophotometric assay kit