Label–free, turn–on fluorescent sensor for trypsin activity assay and inhibitor screening

Zhang, Lufeng; Qin, Haiyan; Cui, Wanwan; Yang, Zhou; Du, Jianxiu

doi:10.1016/j.talanta.2016.09.011

Cited by 22 publications

(11 citation statements)

References 32 publications

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“…S5). The IC 50 value of BH was higher than those reported in the literatures . The difference can be explained by the fact that the experimental conditions were different.…”

Section: Resultscontrasting

confidence: 66%

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Trypsin inhibitor screening in traditional Chinese medicine by using an immobilized enzyme microreactor in capillary and molecular docking study

Cheng

Chen

2017

J of Separation Science

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A trypsin immobilized enzyme microreactor was successfully prepared in capillary for studying enzyme kinetics of trypsin and online screening of trypsin inhibitors from traditional Chinese medicine through capillary electrophoresis. Trypsin was immobilized on the inner wall at the inlet of the capillary treated with polydopamine. The rest of the capillary was used as a separation channel. The parameters including the separation efficiency and the activity of immobilized trypsin were comprehensively evaluated. Under the optimal conditions, online screening of trypsin inhibitors each time can be carried out within 6 min. The Michaelis-Menten constant of immobilized trypsin was calculated to be 0.50 mM, which indicated high affinity of the immobilized trypsin for the substrate. The half-maximal inhibitory concentration of known inhibitor of benzamidine hydrochloride hydrate as a model inhibitor was 13.32 mM. The proposed method was successfully applied to screen trypsin inhibitors from 15 compounds of traditional Chinese medicine. It has been found that baicalin showed inhibitory potency. Molecular docking study well supported the experimental result by exhibiting molecular interaction between enzyme and inhibitors.

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“…S5). The IC 50 value of BH was higher than those reported in the literatures . The difference can be explained by the fact that the experimental conditions were different.…”

Section: Resultscontrasting

confidence: 66%

“…Recently, Zhang et al. developed a fluorescent sensor for trypsin assay by using peroxidase‐like activity of heme‐peptide fragment .…”

Section: Introductionmentioning

confidence: 99%

Trypsin inhibitor screening in traditional Chinese medicine by using an immobilized enzyme microreactor in capillary and molecular docking study

Cheng

Chen

2017

J of Separation Science

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“…After that, the Tryp enzyme solution was dropped on it to immobilize the Tryp on the pores of the NAA. According to previous reports, the Tryp enzyme can cleave Cyt C to the short peptide [ 152 ] fragments, and some of those fragments have a hemin cofactor. The structure of hemin–peptide fragments which are generated by Tryp is as follows: …”

Section: Biosensors (Biorecognizer-based Sensors)mentioning

confidence: 99%

Advances in Optical Biosensors and Sensors Using Nanoporous Anodic Alumina

Tabrizi

Ferré‐Borrull

Marsal

2020

Sensors

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This review paper focuses on recent progress in optical biosensors using self-ordered nanoporous anodic alumina. We present the fabrication of self-ordered nanoporous anodic alumina, surface functionalization, and optical sensor applications. We show that self-ordered nanoporous anodic alumina has good potential for use in the fabrication of antibody-based (immunosensor), aptamer-based (aptasensor), gene-based (genosensor), peptide-based, and enzyme-based optical biosensors. The fabricated optical biosensors presented high sensitivity and selectivity. In addition, we also showed that the performance of the biosensors and the self-ordered nanoporous anodic alumina can be used for assessing biomolecules, heavy ions, and gas molecules.

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“…Current methods for TI screening are all based on activity inhibition assays, including colorimetry , fluorometric assays , and LC combined with biochemical assays . However, these assays need high enzyme consumption, are labor intensive, time consuming and, have low sensitivity sometimes , despite a high level of maturity.…”

Section: Introductionmentioning

confidence: 99%

A strategy for screening trypsin inhibitors from traditional Chinese medicine based on a monolithic capillary immobilized enzyme reactor coupled with offline liquid chromatography and mass spectrometry

Lin

Zhang

Yang

et al. 2019

J of Separation Science

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† These authors contributed equally to this work.A novel strategy was successfully developed for screening trypsin inhibitors in traditional Chinese medicines based on monolithic capillary immobilized enzyme reactors combined with liquid chromatography-tandem mass spectrometry. Organic polymer based monolithic enzyme reactors were firstly prepared by covalently bonding trypsin to a poly(glycidyl methacrylate-co-poly (ethylene glycol) diacrylate) monolith by the ring-opening reaction of epoxy groups. The activity and kinetic parameters of the obtained monolithic trypsin reactors were systematically evaluated using microliquid chromatography. Fourier transform infrared spectroscopy and scanning electron microscopy were also used to characterize the monolithic trypsin reactors. The resulting functional and denatured monolithic trypsin reactors were applied as affinity solidphase extraction columns, and offline coupled with a liquid chromatography-tandem mass spectrometry system to construct a binding affinity screening platform. Subsequently, the proposed platform was applied for screening trypsin binders in a Scutellaria baicalensis Georgi extract. Three compounds, namely scutellarin, baicalin, and wogonoside were identified, and their inhibitory activities were further confirmed via an in vitro enzymatic inhibition assay. Additionally, molecular docking was also performed to study the interactions between trypsin and these three compounds.

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Label–free, turn–on fluorescent sensor for trypsin activity assay and inhibitor screening

Cited by 22 publications

References 32 publications

Trypsin inhibitor screening in traditional Chinese medicine by using an immobilized enzyme microreactor in capillary and molecular docking study

Trypsin inhibitor screening in traditional Chinese medicine by using an immobilized enzyme microreactor in capillary and molecular docking study

Advances in Optical Biosensors and Sensors Using Nanoporous Anodic Alumina

A strategy for screening trypsin inhibitors from traditional Chinese medicine based on a monolithic capillary immobilized enzyme reactor coupled with offline liquid chromatography and mass spectrometry

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