Evaluation of seven cosubstrates in the quantification of horseradish peroxidase enzyme by square wave voltammetry

Kergaravat, Silvina V.; Pividori, María Isabel; Hernández, Silvia R.

doi:10.1016/j.talanta.2011.11.016

Cited by 33 publications

(15 citation statements)

References 33 publications

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“…The lower oxidation potential observed for p-AP in this study compared to the oxidation potential of p-AP on relatively flat gold surfaces reported elsewhere might be related to the nanostructure of NPG[95]. For example, a negative shift in oxidation potential of methanol on NPG has been reported by Zhang et al[96] The peak difference current for SWV has been reported to be proportional to the velocity of the reaction and can be used to create a Michaelis-Menten plot[97, 98]. The rate of a reaction on the electrode surface can be obtained by dividing the peak difference current by the product nF, where, n is the number of electron involved in the redox process and F is Faraday's constant[97].…”

Section: Resultsmentioning

confidence: 68%

Square-wave voltammetry assays for glycoproteins on nanoporous gold

Pandey

Bhattarai

Pornsuriyasak

et al. 2014

Journal of Electroanalytical Chemistry

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Electrochemical enzyme-linked lectinsorbent assays (ELLA) were developed using nanoporous gold (NPG) as a solid support for protein immobilization and as an electrode for the electrochemical determination of the product of the reaction between alkaline phosphatase (ALP) and p-aminophenyl phosphate (p-APP), which is p-aminophenol (p-AP). Glycoproteins or concanavalin A (Con A) and ALP conjugates were covalently immobilized onto lipoic acid self-assembled monolayers on NPG. The binding of Con A – ALP (or soybean agglutinin – ALP) conjugate to glycoproteins covalently immobilized on NPG and subsequent incubation with p-APP substrate was found to result in square-wave voltammograms whose peak difference current varied with the identity of the glycoprotein. NPG presenting covalently bound glycoproteins was used as the basis for a competitive electrochemical assay for glycoproteins in solution (transferrin and IgG). A kinetic ELLA based on steric hindrance of the enzyme-substrate reaction and hence reduced enzymatic reaction rate after glycoprotein binding is demonstrated using immobilized Con A–ALP conjugates. Using the immobilized Con A-ALP conjugate, the binding affinity of immunoglobulin G (IgG) was found to be 105 nM, and that for transferrin was found to be 650 nM. Minimal interference was observed in the presence of 5 mg mL−1 BSA as a model serum protein in both the kinetic and competitive ELLA. Inhibition studies were performed with methyl D-mannoside for the binding of TSF and IgG to Con A-ALP; IC50 values were found to be 90 μM and 286 μM, respectively. Surface coverages of proteins were estimated using solution depletion and the BCA protein concentration assay.

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Section: Resultsmentioning

confidence: 68%

Square-wave voltammetry assays for glycoproteins on nanoporous gold

Pandey

Bhattarai

Pornsuriyasak

et al. 2014

Journal of Electroanalytical Chemistry

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“…Seven phenolic compounds, i.e. the o-phenilendiamine, p-chlorophenol, p-aminophenol hydroquinone, pyrocatechol, phenol and 3,3-5,5-tetramethylbenzidine were tested as co-substrates of the enzymatic reaction of the horseradish peroxidase [124]. The linear dependence between the net SW peak currents and the phenolic substrates concentration was found even in the sub-nanomolar concentration range of studied phenols.…”

Section: Analytical and Biochemical Applications Of Square-wave Voltamentioning

confidence: 99%

Square‐Wave Voltammetry: A Review on the Recent Progress

et al. 2013

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A review on the recent progress of square-wave voltammetry is presented, covering the period of the last five years. The review addresses the new theoretical development of the technique as well as its application for mechanistic purposes, electrode kinetic measurements, biochemical and analytical applications. Besides, a few novel methodological modifications are proposed that might expand the scope and application of the technique.

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“…Our mechanistic model could readily be adapted to other systems and used for other purposes than the ones shown here. For example, Kergaravat et al optimized conditions for electrochemically measuring HRP’s oxidation rate for seven redox-active co-substrates [ 63 ]. For each co-substrate, they optimized the pH and the concentrations of HRP,

, and the co-substrate.…”

Section: Resultsmentioning

confidence: 99%

Integrated Experimental and Theoretical Studies on an Electrochemical Immunosensor

et al. 2020

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Electrochemical immunosensors (EIs) integrate biorecognition molecules (e.g., antibodies) with redox enzymes (e.g., horseradish peroxidase) to combine the advantages of immunoassays (high sensitivity and selectivity) with those of electrochemical biosensors (quantitative electrical signal). However, the complex network of mass-transfer, catalysis, and electrochemical reaction steps that produce the electrical signal makes the design and optimization of EI systems challenging. This paper presents an integrated experimental and modeling framework to address this challenge. The framework includes (1) a mechanistic mathematical model that describes the rate of key mass-transfer and reaction steps; (2) a statistical-design-of-experiments study to optimize operating conditions and validate the mechanistic model; and (3) a novel dimensional analysis to assess the degree to which individual mass-transfer and reaction steps limit the EI’s signal amplitude and sensitivity. The validated mechanistic model was able to predict the effect of four independent variables (working electrode overpotential, pH, and concentrations of catechol and hydrogen peroxide) on the EI’s signal magnitude. The model was then used to calculate dimensionless groups, including Damkohler numbers, novel current-control coefficients, and sensitivity-control coefficients that indicated the extent to which the individual mass-transfer or reaction steps limited the EI’s signal amplitude and sensitivity.

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Evaluation of seven cosubstrates in the quantification of horseradish peroxidase enzyme by square wave voltammetry

Cited by 33 publications

References 33 publications

Square-wave voltammetry assays for glycoproteins on nanoporous gold

Square-wave voltammetry assays for glycoproteins on nanoporous gold

Square‐Wave Voltammetry: A Review on the Recent Progress

Integrated Experimental and Theoretical Studies on an Electrochemical Immunosensor

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