A simple fluorescent probe based on a pyrene derivative for rapid detection of protamine and monitoring of trypsin activity

Tang, Baiyang; Yang, Yan; Wang, Gefu; Yao, Zhiyi; Zhang, Li; Wu, Hai‐Chen

doi:10.1039/c5ob01034a

Cited by 35 publications

(13 citation statements)

References 44 publications

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“…Trypsin can be determined with a linear range of 0.0–50.0 ng/mL (inset in Figure 3C). The detection limit of trypsin was calculated as 1 ng/mL according to the rule of three times the standard deviation over the blank response, which was lower than that of some fluorescence assays reported recently [9,12,13,41,42,43]. The label-free properties of the method can offer the advantages of easy preparation of DNA-AgNCs, low cost, and operation convenience.…”

Section: Resultsmentioning

confidence: 81%

Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

Zhuo

Wang

Feng

et al. 2016

Sensors

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Trypsin is important during the regulation of pancreatic exocrine function. The detection of trypsin activity is currently limited because of the need for the substrate to be labeled with a fluorescent tag. A label-free fluorescent method has been developed to monitor trypsin activity. The designed peptide probe consists of six arginine molecules and a cysteine terminus and can be conjugated to DNA-stabilized silver nanoclusters (DNA-AgNCs) by Ag-S bonding to enhance fluorescence. The peptide probe can also be adsorbed to the surface of graphene oxide (GO), thus resulting in the fluorescence quenching of DNA-AgNCs-peptide conjugate because of Förster resonance energy transfer. Once trypsin had degraded the peptide probe into amino acid residues, the DNA-AgNCs were released from the surface of GO, and the enhanced fluorescence of DNA-AgNCs was restored. Trypsin can be determined with a linear range of 0.0–50.0 ng/mL with a concentration as low as 1 ng/mL. This label-free method is simple and sensitive and has been successfully used for the determination of trypsin in serum. The method can also be modified to detect other proteases.

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Section: Resultsmentioning

confidence: 81%

Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

Zhuo

Wang

Feng

et al. 2016

Sensors

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“…It is well known that certain carbon nanomaterials, including graphene, fullerene, carbon nanotubes, and carbon nanofibers, among others, have attracted much attention at the frontiers of technological innovation as drug delivery carriers due to their excellent electrochemical, optical, mechanical biocompatibility, thermal and adsorption properties [ [10] , [11] , [12] ]. Nevertheless, there are relatively few studies regarding carbon nanoparticle materials compared to the above carbon-based materials.…”

Section: Introductionmentioning

confidence: 99%

A carbon nanoparticle-peptide fluorescent sensor custom-made for simple and sensitive detection of trypsin

Hou

Feng

Zhao

et al. 2020

Journal of Pharmaceutical Analysis

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Herein, we report a novel sensor to detect trypsin using a purpose-designed fluorescein-labelled peptide with negatively charged carbon nanoparticles (CNPs) modified by acid oxidation. The fluorescence of the fluorescein-labelled peptide was quenched by CNPs. The sensor reacted with trypsin to cleave the peptide, resulting in the release of the dye moiety and a substantial increase in fluorescence intensity, which was dose- and time-dependent, and trypsin could be quantified accordingly. Correspondingly, the biosensor has led to the development of a convenient and efficient fluorescent method to measure trypsin activity, with a detection limit of 0.7 μg/mL. The method allows rapid determination of trypsin activity in the normal and acute pancreatitis range, suitable for point-of-care testing. Furthermore, the applicability of the method has been demonstrated by detecting trypsin in spiked urine samples.

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“…Protamine is a class of polycationic proteins, where approximately 67% of the contents are contributed by strongly basic arginine residues. , It has a molecular weight of around 4.5 kDa, and its isoelectric point is at 13.8. , Protamine contributes significantly in many biological mechanisms, including hematopoiesis and bone metabolism . With its high positive charge density and the strong binding affinity with heparin, protamine, which can bind to anionic heparin electrostatically, is regarded as an antidote to neutralize the anticoagulant effect from heparin .…”

Section: Introductionmentioning

confidence: 99%

Protamine-Induced Supramolecular Self-Assembly of Red-Emissive Alkynylplatinum(II) 2,6-Bis(benzimidazol-2′-yl)pyridine Complex for Selective Label-Free Sensing of Heparin and Real-Time Monitoring of Trypsin Activity

Chan

Cheng

Hau

et al. 2019

ACS Appl. Mater. Interfaces

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A label-free detection assay is developed based on the design and synthesis of a new anionic alkynylplatinum(II) 2,6-bis(benzimidazol-2′-yl)pyridine complex with water-soluble pendants. With the aid of electrostatic interaction and noncovalent metal–metal and π–π stacking interactions, protamine is shown to induce supramolecular self-assembly of platinum(II) complexes with drastic UV–vis absorption and red emission changes. On the basis of the strong binding affinity of protamine and heparin, the ensemble has been further employed to probe heparin by monitoring the spectroscopic changes. Other than heparin, this ensemble can also detect the activity of trypsin, which can hydrolyze protamine into fragments, leading to the deaggregation of platinum(II) complexes. By modulation of the self-assembly properties of platinum(II) complexes via real-time UV–vis absorption and emission studies, the reported assay has been demonstrated to be a sensitive and selective detection method for trypsin, as well as trypsin inhibitor screening, which is essential for drug discovery.

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A simple fluorescent probe based on a pyrene derivative for rapid detection of protamine and monitoring of trypsin activity

Cited by 35 publications

References 44 publications

Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

Label-Free Fluorescent Detection of Trypsin Activity Based on DNA-Stabilized Silver Nanocluster-Peptide Conjugates

A carbon nanoparticle-peptide fluorescent sensor custom-made for simple and sensitive detection of trypsin

Protamine-Induced Supramolecular Self-Assembly of Red-Emissive Alkynylplatinum(II) 2,6-Bis(benzimidazol-2′-yl)pyridine Complex for Selective Label-Free Sensing of Heparin and Real-Time Monitoring of Trypsin Activity

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