Usefulness of magnetically-controlled MNPs-enzymes microreactors for the fluorimetric determination of total cholesterol in serum

Román-Pizarro, Vanesa; Ramírez-Gutiérrez, Manuel; Gómez‐Hens, A.; Fernández-Romero, J.M.

doi:10.1016/j.talanta.2019.120426

Cited by 10 publications

(9 citation statements)

References 25 publications

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“…Its dynamic level in blood is a significant biomarker for estimating the patient conditions and predicting the possibility of shock, collapse, and mortality. , Cholesterol has a vital role in maintaining the structure and function of the cell membrane. The imbalance of its concentration in the human body is often associated with some diseases, such as Alzheimer’s disease, hypertension, coronary heart disease, and myocardial infarction. − Because of the significance of these biological substances and their coexistence in the human body, a precise and sensitive method enabling their simultaneous quantitative detection is highly needed. , …”

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confidence: 99%

Reusable OIRD Microarray Chips Based on a Bienzyme-Immobilized Polyaniline Nanowire Forest for Multiplexed Detection of Biological Small Molecules

et al. 2021

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Quantitative detection of multiple biological small molecules is critical for health evaluation and disease diagnosis. In this study, a microarray chip featuring a bienzyme-immobilized polyaniline nanowire forest on fluorine-doped tin oxide (bienzyme-PANI/FTO) is developed for this purpose. On such a chip, the target molecules are oxidized under the catalysis of their attached oxidases to produce hydrogen peroxide, which further induces the partial oxidation of local PANI nanowires in the presence of horseradish peroxidase (HRP) enzyme. The redox state change of PANI nanowires is monitored by the oblique incident reflectivity difference (OIRD) technique in a real-time and wireless manner, thus allowing for quantitative analysis of the target molecules. As typical model targets, hydrogen peroxide, glucose, lactic acid, and cholesterol are successfully detected with low detection limits, excellent specificities, and broad detection ranges, all of which fully meet the requirements for clinical analysis of human serum samples. Simultaneous detection of multiple targets on an individual chip is further demonstrated using the OIRD scanning mode. Meanwhile, by simple electrochemical reduction of the PANI nanowires, the chip is reusable for more than eight detection cycles without evident decay in its performance. The detection principle of this chip is also universal to other small molecules, and thus, it shows great promise as a valuable device to analyze biological small molecules.

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confidence: 99%

Reusable OIRD Microarray Chips Based on a Bienzyme-Immobilized Polyaniline Nanowire Forest for Multiplexed Detection of Biological Small Molecules

et al. 2021

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“…22,23 For instance, cholesterol determination was performed in a tubular device with co-immobilized cholesterol esterase/cholesterol oxidase-MNPs affixed to a permanent magnet. 20 MNP-bound pectinase acted with much higher biocatalytic efficiency in an MNP-based membrane reactor than in a batch reactor. 21 MNPs proved to be a suitable support material for immobilization of phenylalanine ammonia-lyase (PAL) as a biocatalyst (PAL-MNP) for biotransformations in microfluidic systems.…”

Section: Introductionmentioning

confidence: 95%

Magnetically agitated continuous-flow tube reactors with aspartate ammonia-lyase immobilized on magnetic nanoparticles

et al. 2023

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“…Before their use for enzyme immobilization, the solvent was evaporated until completely dried in an oven at 90 °C until the weight remained constant in five successive weightings. The immobilization of the GPx enzyme was carried out by a covalent binding using the cross-linking carbodiimide reaction, which modified the immobilization of other enzymes [8,33,34]. Initially, 0.0498 g of dried MNPs were added to 1 mL of phosphate buffer (50 mmol L −1 , pH 7.4), and the solution was sonicated for 15 min after adding 0.5 mL of a 0.02 g mL −1 EDC solution dissolved in the phosphate buffer mentioned.…”

Section: Synthesis Of Gpx-mnps Complexmentioning

confidence: 99%

“…The characterization of the GPx-MNPs complex has been avoided because MNPs and the complex with enzymes have been previously described [32,34]. After the immobilization of the enzyme, "batch" tests were performed to evaluate the efficiency of the GPx immobilization process on MNPs.…”

Section: Synthesis Of Gpx-mnps Complexmentioning

confidence: 99%

Study of the inhibition effects on glutathione peroxidase immobilized on MNPs using a stopped-flow microfluidic system

et al. 2023

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A stopped-flow microfluidic system to monitor glutathione peroxidase (GPx) activity and evaluate potential inhibitors of the enzyme has been developed based on the integration of the microfluidic chip in the reaction/detection zone. This integration supposes the physical alignment at the optimal location of the microfluidic channel, both the magnetically retained enzyme microreactor (MREµR) and the remote luminescence detection using a focused bifurcated fiber optic bundle (BFOB) connected to a conventional spectrofluorometer detector. The method is based on the coupling of two competitive oxidative chemical reactions, in which glutathione (GSH) and homovanillic acid (HVA) competed for their interaction with hydrogen peroxide in the presence of the magnetically retained GPx-MNPs. The biocatalytic reaction was followed by monitoring the fluorescence of the biphenyl-HVA dimer formed. The dynamic range of the calibration graph was 0.45–10 µmol L−1, expressed as GSH concentration with a detection limit of 0.1 µmol L−1 (r2 = 0.9954, n = 10, r = 3). The precision expressed as the relative standard deviation (RSD%) was between 0.5 and 3.9%. The stopped-flow microfluidic system showed a sampling frequency of 25 h−1. The method was applied to the study of GPx inhibition provided by three inhibitory compounds, two metallic ions Hg(II) and Cu(II) and t-butyl hydroperoxide, and their presence in liquid samples, as water, milk, and edible oil. Recovery values between 88.7 and 99.4% were achieved in all instances. Graphical Abstract

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Usefulness of magnetically-controlled MNPs-enzymes microreactors for the fluorimetric determination of total cholesterol in serum

Cited by 10 publications

References 25 publications

Reusable OIRD Microarray Chips Based on a Bienzyme-Immobilized Polyaniline Nanowire Forest for Multiplexed Detection of Biological Small Molecules

Reusable OIRD Microarray Chips Based on a Bienzyme-Immobilized Polyaniline Nanowire Forest for Multiplexed Detection of Biological Small Molecules

Magnetically agitated continuous-flow tube reactors with aspartate ammonia-lyase immobilized on magnetic nanoparticles

Study of the inhibition effects on glutathione peroxidase immobilized on MNPs using a stopped-flow microfluidic system

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