Advances in screening enzyme inhibitors by capillary electrophoresis

Wang, Weifeng; Yang, Jun‐Li

doi:10.1002/elps.201900013

Cited by 21 publications

(17 citation statements)

References 79 publications

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“…Enzyme inhibitor assay is an essential tool for drug discovery. 1 It usually involves the analysis of a significant quantity of bioactive compounds in large libraries. Therefore, time-saving, high-throughput, and simple techniques using low sample volumes are required.…”

Section: Introductionmentioning

confidence: 99%

“…Therefore, time-saving, high-throughput, and simple techniques using low sample volumes are required. [1][2][3] To achieve these objectives, several enzyme inhibitor assay-related approaches have been reported. [4][5][6][7][8][9][10][11][12][13] In enzyme inhibitor assays, high-performance liquid chromatography (HPLC) with ultraviolet (UV) detection, and capillary electrophoresis (CE) are widely used as analytical methods.…”

Section: Introductionmentioning

confidence: 99%

“…However, electrophoretic separation-based CE allows high-efficiency separation; multiple enzyme inhibitor assay methods using CE have thus been reported. 1,3,14,[18][19][20][21] However, CE requires large equipment and involves multi-step reactions.…”

Section: Introductionmentioning

confidence: 99%

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Single-step Trypsin Inhibitor Assay on a Microchannel Array Device Immobilizing Enzymes and Fluorescent Substrates by Inkjet Printing

et al. 2021

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In this study, we report a single-step trypsin inhibitor assay on a microchannel array device immobilizing enzymes and substrates by inkjet printing. The microdevice is composed of a poly(dimethylsiloxane) (PDMS) microchannel array that immobilizes trypsin and fluorescent substrates as reactive reagents at the two bottom corners of a microchannel. Inkjet printers allow simple, accurate, and position-selective immobilization of reagents as nanoliter spots. Therefore, plural reactive reagents, such as enzymes and substrates, can be separately immobilized at different positions in the same microchannel without mixing, allowing single-step operation by simply introducing a sample solution through capillary action. Furthermore, reproducible fabrication and mass production of the device could be expected. In this study, the efficiency of an acidic solution as a spotting agent for protease immobilization to prevent the decrease in fluorescence intensity was confirmed. Additionally, single-step trypsin inhibitor screening was performed using three inhibitors. Finally, we investigated the storage stability of the device and confirmed that it remained stable for at least 10 days.

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Section: Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Single-step Trypsin Inhibitor Assay on a Microchannel Array Device Immobilizing Enzymes and Fluorescent Substrates by Inkjet Printing

et al. 2021

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“…Up to date, many modern analysis instruments, including UV‐vis spectrum, fluorescence spectrophotometry, radioisotope labeling, HPLC, electrochemical analysis, and CE, have been explored to conquer the disadvantage of lower efficiency, narrow application range, environmental pollution and long analysis time, for drug research and enzyme inhibitor screening [11–15]. Due to the advantages of high separation efficiency, fast analysis speed and low sample consumption, CE is very suitable for the microanalysis of rare and complex samples, and has been widely used in the field of biomolecule, enzyme activity evaluation, and drug discovery [16–19]. Since Banke et al.…”

Section: Introductionmentioning

confidence: 99%

An online target and rapid screening method for α‐glucosidase inhibitors based on capillary electrophoresis

et al. 2021

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Screening enzymatic active compounds is one of the important fields in drug research. α‐Glucosidase can hydrolyze carbohydrates to monosaccharides after meals and lead to the rise of blood glucose levels in human body. Thus, the inhibition of α‐glucosidase activity is an effective approach for the diabetes treatment. In this work, we developed a new method to simultaneously screen multiple bioactive compounds within a single CE running. The affect factors on the method performance, including injection, mixing, incubation, separation and detection, were carefully analyzed and discussed. Under the optimum, the mixture consisting of two internal standards (DMSO and 4‐nitrophenol) and five compounds (lyoniresinol, hydroxytyrosol, rutin, kaempferol, and quercetin) was simultaneously screened, and kaempferol and quercetin showed stronger activity and this conclusion was also supported by offline assay. Furthermore, molecular docking was employed for investigating its interaction mechanism. Eventually, the established method has been applied to screen potential α‐glucosidase inhibitors from an extract of Lycium barbarum and the peak area of rutin, taxifolin, quercetin, and chlorogenic acid in L. barbarum samples changed before and after the enzymatic reaction, confirming that these four compounds had potential inhibitory activities, which was consistent with the literature data. The present work provides a promising method for the target and rapid discovery of bioactive compounds from a plant extract or mixture.

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“…However, the post-column derivation made the operation more complicated. The capillary electrophoresis (CE) has the advantages of low solvent consumption and high efficiency, so it was widely applied in screening enzyme inhibitors, complex samples analysis, and derivatization [18,19,20] in recent years. At present, the in-capillary ABTS + -CE-DAD method [21] and DPPH-CE-DAD method [22,23] were established to evaluate the antioxidant activity of TCM.…”

Section: Introductionmentioning

confidence: 99%

Microwave-Assisted Extraction Combined with In-Capillary [Fe(ferrozine)3]2+-CE-DAD to Screen Active Components with the Ability to Chelate Ferrous Ions from Flos Sophorae Immaturus (Flos Sophorae)

Liu

Wang

et al. 2019

Molecules

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An efficient microwave-assisted extraction (MAE) combined with in-capillary [Fe(ferrozine)3]2+-capillary electrophoresis-Diode Array Detector (in-capillary [Fe(ferrozine)3]2+-CE-DAD) was developed to screen active components with the ability to chelate ferrous ions and determine the total antioxidant activity. The MAE conditions, including methanol concentration, extraction power, extraction time, and the ratio of material to liquid, were optimized by an L9(34) orthogonal experiment. Background buffer, voltage, and cartridge temperature that affect the separation of six compounds were optimized. It was found that rutin and quercetin were the main components chelating ferrous ions in Flos Sophorae Immaturus (Flos Sophorae) by the in-capillary [Fe(ferrozine)3]2+-CE-DAD. The recoveries were ranged from 95.2% to 104%. It was concluded that the MAE combined with in-capillary [Fe(ferrozine)3]2+-CE-DAD method was a simple, reliable, and efficient tool for screening active components from the complex traditional Chinese medicine samples and evaluating their ability to chelate ferrous ions.

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Advances in screening enzyme inhibitors by capillary electrophoresis

Cited by 21 publications

References 79 publications

Single-step Trypsin Inhibitor Assay on a Microchannel Array Device Immobilizing Enzymes and Fluorescent Substrates by Inkjet Printing

Single-step Trypsin Inhibitor Assay on a Microchannel Array Device Immobilizing Enzymes and Fluorescent Substrates by Inkjet Printing

An online target and rapid screening method for α‐glucosidase inhibitors based on capillary electrophoresis

Microwave-Assisted Extraction Combined with In-Capillary [Fe(ferrozine)3]2+-CE-DAD to Screen Active Components with the Ability to Chelate Ferrous Ions from Flos Sophorae Immaturus (Flos Sophorae)

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