Lactate biosensing based on covalent immobilization of lactate oxidase onto chevron-like graphene nanoribbons via diazotization-coupling reaction
Entity
UAM. Departamento de Química Analítica y Análisis InstrumentalPublisher
ElsevierDate
2022-04-20Citation
10.1016/j.aca.2022.339851
Analytica Chimica Acta 1208 (2022): 339851
ISSN
0003-2670 (print)DOI
10.1016/j.aca.2022.339851Funded by
The authors acknowledge financial support from the European Union’s Horizon 2020 research and innovation programme (Project SPRING, grant agreement No 863098), the European Research Council (ERC) (Project MolDAM, grant agreement No 951519), Ministerio de Ciencia e Innovacion ´ of Spain (Grants MAT2017-85089-C2-1-R, MAT2017-85089-C2-2-R, PID2020-113142RB-C21, PID2020- 113142RB-C22) funded by MCIN/AEI/10.13039/501100011033, the Comunidad Autonoma ´ de Madrid (S2018/NMT-4349TRANSNANOAVANSENS-CM), the Xunta de Galicia (Centro singular de investigacion ´ de Galicia accreditation 2019–2022, ED431G 2019/03) and the European Union (European Regional Development Fund - ERDF). J. Castro thanks the Spanish MINECO and the European Social Fund for the predoctoral grant BES-2017-081094. We want to thank Ismael Ballesteros for the SEM measurementsProject
info:eu-repo/grantAgrement/EC/H2020/863098/EU//SPRING; info:eu-repo/grantAgreement/EC/H2020/951519/EU//MolDAM; Gobierno de España. PID2019-106188GB-I00Editor's Version
https://doi.org/10.1016/j.aca.2022.339851Subjects
Biosensor; Chevron-like graphene nanoribbons; Lactate oxidase; Diazonium chemistry; Electrochemical detection; Lactate; QuímicaRights
© 2022 The AuthorsEsta obra está bajo una licencia de Creative Commons Reconocimiento-NoComercial-SinObraDerivada 4.0 Internacional.
Abstract
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
Files in this item
Google Scholar:Sainz Vaque, Raquel
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Pozo Vázquez, María del
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Vazquez, Luís
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Vilas Varela, Manuel
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Castro Esteban, Jesús
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Blanco Gil, Elías
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Petit Domínguez, María Dolores
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Quintana Mani, María del Carmen
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Casero Junquera, María Elena
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