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Le DV. Fluorescence Determination of Peptidase Activity Based on the Quenching of a Fluorophore-Labelled Peptide by Graphene Oxide. Protein J. 2021;40(5):682-688doi: 10.1007/s10930-021-09985-6.
Le, D. V. (2021). Fluorescence Determination of Peptidase Activity Based on the Quenching of a Fluorophore-Labelled Peptide by Graphene Oxide. The protein journal, 40(5), 682-688. https://doi.org/10.1007/s10930-021-09985-6
Le, Dinh-Vu. "Fluorescence Determination of Peptidase Activity Based on the Quenching of a Fluorophore-Labelled Peptide by Graphene Oxide." The protein journal vol. 40,5 (2021): 682-688. doi: https://doi.org/10.1007/s10930-021-09985-6
Le DV. Fluorescence Determination of Peptidase Activity Based on the Quenching of a Fluorophore-Labelled Peptide by Graphene Oxide. Protein J. 2021 Oct;40(5):682-688. doi: 10.1007/s10930-021-09985-6. Epub 2021 Apr 15. PMID: 33856620.
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Protein J. 2021 Oct;40(5):682-688. doi: 10.1007/s10930-021-09985-6. Epub 2021 Apr 15.

Fluorescence Determination of Peptidase Activity Based on the Quenching of a Fluorophore-Labelled Peptide by Graphene Oxide.

The protein journal

Dinh-Vu Le

Affiliations

  1. Faculty of Chemical Engineering, Industrial University of Ho Chi Minh City, 12 Nguyen Van Bao St. Go Vap, Ho Chi Minh City, 70000, Vietnam. [email protected].

PMID: 33856620 DOI: 10.1007/s10930-021-09985-6

Abstract

In this study, a fluorescence detection strategy is reported for the peptidase activity assay, which is based on fluorescence resonance energy transfer (FRET) from a fluorophore-labelled peptide to graphene oxide (GO). By the hydrolysis of the peptide, the fluorophore-labelled peptide releases the fluorophore 5-carboxyfluorescein, which can avoid quenching from GO. Thus, the increased intensity of the obtained fluorescence signal in the assay is directly dependent on the peptidase activity. As a model case of the developed strategy, the activity determination of pancreatic elastase (PE) is performed. Under the optimal experimental conditions at an excitation wavelength of 494 nm, the activity of PE can be determined in the range from 0.003 to 0.10 U/mL, with a detection limit of 0.001 U/mL at the emission wavelength of 518 nm. This is ultra-sensitive for the determination of PE. The specificity of the method is demonstrated by the analysis of PE under complex conditions using fetal bovine serum as the substrate. Hence, the developed method might provide an intrinsically convenient, sensitive platform for the PE activity assay and related biochemical studies due to its homogeneous, and fluorescence-based detection strategy.

© 2021. The Author(s), under exclusive licence to Springer Science+Business Media, LLC, part of Springer Nature.

Keywords: Fluorescence quenching; Graphene oxide; Pancreatic elastase; Peptidase

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