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1.
Mikrochim Acta ; 191(9): 558, 2024 08 23.
Artículo en Inglés | MEDLINE | ID: mdl-39177820

RESUMEN

An innovative supramolecular architecture is reported for bienzymatic glucose biosensing based on the use of a nanohybrid made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with a Schiff base modified with two phenylboronic acid residues (SB-dBA) as platform for the site-specific immobilization of the glycoproteins glucose oxidase (GOx) and horseradish peroxidase (HRP). The analytical signal was obtained from amperometric experiments at - 0.050 V in the presence of 5.0 × 10-4 M hydroquinone as redox mediator. The concentration of GOx and HRP and the interaction time between the enzymes and the nanohybrid MWCNT-SB-dBA deposited at glassy carbon electrodes (GCEs) were optimized through a central composite design (CCD)/response surface methodology (RSM). The optimal concentrations of GOx and HRP were 3.0 mg mL-1 and 1.50 mg mL-1, respectively, while the optimum interaction time was 3.0 min. The bienzymatic biosensor presented a sensitivity of (24 ± 2) × 102 µA dL mg-1 ((44 ± 4) × 102 µA M-1), a linear range between 0.06 mg dL-1 and 21.6 mg dL-1 (3.1 µM-1.2 mM) (R2 = 0.9991), and detection and quantification limits of 0.02 mg dL-1 (1.0 µM) and 0.06 mg dL-1 (3.1 µM), respectively. The reproducibility for five sensors prepared with the same MWCNT-SB-dBA nanohybrid was 6.3%, while the reproducibility for sensors prepared with five different nanohybrids and five electrodes each was 7.9%. The GCE/MWCNT-SB-dBA/GOx-HRP was successfully used for the quantification of glucose in artificial human urine and commercial human serum samples.


Asunto(s)
Técnicas Biosensibles , Ácidos Borónicos , Enzimas Inmovilizadas , Glucosa Oxidasa , Peroxidasa de Rábano Silvestre , Nanotubos de Carbono , Bases de Schiff , Nanotubos de Carbono/química , Bases de Schiff/química , Técnicas Biosensibles/métodos , Ácidos Borónicos/química , Glucosa Oxidasa/química , Glucosa Oxidasa/metabolismo , Peroxidasa de Rábano Silvestre/química , Peroxidasa de Rábano Silvestre/metabolismo , Humanos , Enzimas Inmovilizadas/química , Enzimas Inmovilizadas/metabolismo , Glucosa/análisis , Electrodos , Límite de Detección , Técnicas Electroquímicas/métodos , Glucemia/análisis
2.
Biosensors (Basel) ; 14(4)2024 Apr 16.
Artículo en Inglés | MEDLINE | ID: mdl-38667189

RESUMEN

L-Lactate is an important bioanalyte in the food industry, biotechnology, and human healthcare. In this work, we report the development of a new L-lactate electrochemical biosensor based on the use of multiwalled carbon nanotubes non-covalently functionalized with avidin (MWCNT-Av) deposited at glassy carbon electrodes (GCEs) as anchoring sites for the bioaffinity-based immobilization of a new recombinant biotinylated lactate oxidase (bLOx) produced in Escherichia coli through in vivo biotinylation. The specific binding of MWCNT-Av to bLOx was characterized by amperometry, surface plasmon resonance (SPR), and electrochemical impedance spectroscopy (EIS). The amperometric detection of L-lactate was performed at -0.100 V, with a linear range between 100 and 700 µM, a detection limit of 33 µM, and a quantification limit of 100 µM. The proposed biosensor (GCE/MWCNT-Av/bLOx) showed a reproducibility of 6.0% and it was successfully used for determining L-lactate in food and enriched serum samples.


Asunto(s)
Avidina , Técnicas Biosensibles , Ácido Láctico , Oxigenasas de Función Mixta , Nanotubos de Carbono , Nanotubos de Carbono/química , Oxigenasas de Función Mixta/química , Avidina/química , Técnicas Electroquímicas , Resonancia por Plasmón de Superficie , Enzimas Inmovilizadas/química , Escherichia coli , Biotinilación , Electrodos , Espectroscopía Dieléctrica , Límite de Detección
3.
Mikrochim Acta ; 190(2): 73, 2023 01 25.
Artículo en Inglés | MEDLINE | ID: mdl-36695940

RESUMEN

An innovative strategy is proposed to simultaneously exfoliate multi-walled carbon nanotubes (MWCNTs) and generate MWCNTs with immunoaffinity properties. This strategy was based on the non-covalent functionalization of MWCNTs with human immunoglobulin G (IgG) by sonicating 2.5 mg mL-1 MWCNTs in 2.0 mg mL-1 IgG for 15 min with sonicator bath. Impedimetric experiments performed at glassy carbon electrodes (GCE) modified with the resulting MWCNT-IgG nanohybrid in the presence of anti-human immunoglobulin G antibody (Anti-IgG) demonstrated that the immunoglobulin retains their biorecognition properties even after the treatment during the MWCNT functionalization. We proposed, as proof-of-concept, two model electrochemical sensors, a voltammetric one for uric acid quantification by taking advantages of the exfoliated MWCNTs electroactivity (linear range, 5.0 × 10-7 M - 5.0 × 10-6 M; detection limit, 165 nM) and an impedimetric immunosensor for the detection of Anti-IgG through the use of the bioaffinity properties of the IgG present in the nanohybrid (linear range, 5-50 µg mL-1; detection limit, 2 µg mL-1).


Asunto(s)
Técnicas Biosensibles , Nanotubos de Carbono , Humanos , Técnicas Biosensibles/métodos , Nanotubos de Carbono/química , Inmunoensayo , Inmunoglobulina G , Electrodos
4.
Biosens Bioelectron X ; 12: 100222, 2022 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-36118917

RESUMEN

We report two novel genosensors for the quantification of SARS-CoV-2 nucleic acid using glassy carbon electrodes modified with a biocapture nanoplatform made of multi-walled carbon nanotubes (MWCNTs) non-covalently functionalized with avidin (Av) as a support of the biotinylated-DNA probes. One of the genosensors was based on impedimetric transduction offering a non-labelled and non-amplified detection of SARS-CoV-2 nucleic acid through the increment of [Fe(CN)6]3-/4- charge transfer resistance. This biosensor presented an excellent analytical performance, with a linear range of 1.0 × 10-18 M - 1.0 × 10-11 M, a sensitivity of (5.8 ± 0.6) x 102 Ω M-1 (r2 = 0.994), detection and quantification limits of 0.33 aM and 1.0 aM, respectively; and reproducibilities of 5.4% for 1.0 × 10-15 M target using the same MWCNTs-Av-bDNAp nanoplatform, and 6.9% for 1.0 × 10-15 M target using 3 different nanoplatforms. The other genosensor was based on a sandwich hybridization scheme and amperometric transduction using the streptavidin(Strep)-biotinylated horseradish peroxidase (bHRP)/hydrogen peroxide/hydroquinone (HQ) system. This genosensor allowed an extremely sensitive quantification of the SARS-CoV-2 nucleic acid, with a linear range of 1.0 × 10-20 M - 1.0 × 10-17 M, detection limit at zM level, and a reproducibility of 11% for genosensors prepared with the same MWCNTs-Av-bDNAp1 nanoplatform. As a proof-of-concept, and considering the extremely high sensitivity, the genosensor was challenged with highly diluted samples obtained from SARS-CoV-2 RNA PCR amplification.

5.
Analyst ; 147(10): 2130-2140, 2022 May 17.
Artículo en Inglés | MEDLINE | ID: mdl-35421882

RESUMEN

This work presents for the first time the systematic preparation of a novel carbon nanotube-MCM-41 hybrid employing the mesoporous material MCM-41 as a successful dispersant for multiwall carbon nanotubes (MWCNTs). Relevant dispersion variables such as the amount of MWCNTs, MCM-41 concentration, and sonication time were optimized through a central composite design (CDD)/response surface methodology (RSM). Several solvents were evaluated and N,N-dimethylformamide (DMF) was selected because it allowed reaching stable dispersions with very good electrochemical response. The electrochemical performance of glassy carbon electrodes (GCE) modified with different hybrids was evaluated by cyclic voltammetry (CV) using ascorbic acid (AA) as redox marker, while their surface morphology was characterized by SEM microscopy. The optimal MWCNT-MCM-41 dispersion condition was 0.75 mg mL-1 MWCNTs, 0.25 mg mL-1 MCM-41, and 30 min sonication. Both, electrochemical results and SEM images correlate with a percolation behavior from MWCNT-MCM-41 hybrid. Electrooxidation of AA at GCE modified with the optimal hybrid occurred under diffusion control and exhibited an enhanced current response (65 µA) and a lower overvoltage (-0.005 V) compared to bare GCE (ip = 22 µA, Ep = 0.255 V). The amperometric response of AA at GCE/MWCNT-MCM-41 exhibited remarkable figures of merit, including an ultralow detection limit (1.5 nM), high sensitivity (45.4 × 103 µA M-1), excellent short- and long-term stability, and very good anti-interference ability for AA detection. The analytical applicability of the developed electrochemical sensor was evaluated by sensing AA in several real samples, showing excellent correlation with the values reported by manufacturers in both pharmaceutical and food samples.


Asunto(s)
Nanotubos de Carbono , Ácido Ascórbico/química , Técnicas Electroquímicas/métodos , Electrodos , Nanotubos de Carbono/química , Dióxido de Silicio
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