Protein Encapsulation: A Nanocarrier Approach to the Fluorescence Imaging of an Enzyme-Based Biomarker

Zhiyuan Jia; Hai Hao Han; Adam C. Sedgwick; George T. Williams; Lauren Gwynne; James T. Brewster; Steven D. Bull; A. Toby A. Jenkins; Xiao Peng He; Holger Schönherr; Jonathan L. Sessler; Tony D. James

doi:10.3389/fchem.2020.00389

Protein Encapsulation: A Nanocarrier Approach to the Fluorescence Imaging of an Enzyme-Based Biomarker

Zhiyuan Jia, Hai Hao Han, Adam C. Sedgwick, George T. Williams, Lauren Gwynne, James T. Brewster, Steven D. Bull, A. Toby A. Jenkins, Xiao Peng He, Holger Schönherr, Jonathan L. Sessler, Tony D. James

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24 Citations (SciVal)

Abstract

Here, we report a new pentafluoropropanamido rhodamine fluorescent probe (ACS-HNE) that allows for the selective detection of neutrophil elastase (NE). ACS-HNE displayed high sensitivity, with a low limit of detection (<5.3 nM), and excellent selectivity toward elastase over other relevant biological analytes and enzymes. The comparatively poor solubility and cell permeability of neat ACS-HNE was improved by creating an ACS-HNE-albumin complex; this approach allowed for improvements in the in situ visualization of elastase activity in RAW 264.7 cells relative to ACS-HNE alone. The present study thus serves to demonstrate a simple universal strategy that may be used to overcome cell impermeability and solubility limitations, and to prepare probes suitable for the cellular imaging of enzymatic activity in vitro.

Original language	English
Article number	389
Journal	Frontiers in Chemistry
Volume	8
DOIs	https://doi.org/10.3389/fchem.2020.00389
Publication status	Published - 3 Jun 2020

Funding

AS thanks the EPSRC for a studentship. TJ wishes to thank the Royal Society for a Wolfson Research Merit Award. NMR characterization facilities were provided through the Chemical Characterization and Analysis Facility (CCAF) at the University of Bath (www.bath.ac.uk/ccaf). The EPSRC UK National Mass Spectrometry Facility at Swansea University is thanked for analyses. Funding. We would like to thank the EPSRC and the University of Bath for funding. We are grateful for financial support by the European Research Council (ERC grant to HS, ERC grant agreement No. 279202) and the University of Siegen. The work in Austin was supported by the National Institutes of Health (RO1 GM103790 to JS) and the Robert A. Welch Foundation (F-0018 to JS). The work at ECUST was supported by the National Natural Science Foundation of China (Nos. 21788102, 91853201 and 21722801), the Shanghai Municipal Science and Technology Major Project (No. 2018SHZDZX03), the International Cooperation Program of Shanghai Science and Technology Committee (No. 17520750100) and the Fundamental Research Funds for the Central Universities (222201717003). This work was supported in part by grant MR/N0137941/1 for the GW4 BIOMED DTP, awarded to the Universities of Bath, Bristol, Cardiff, and Exeter from the Medical Research Council (MRC)/UKRI.

Keywords

BSA-based nanocarrier
cell imaging
elastase detection
fluorescence imaging
nanocarrier-based enzyme detection

ASJC Scopus subject areas

General Chemistry

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10.3389/fchem.2020.00389Licence: CC BY

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title = "Protein Encapsulation: A Nanocarrier Approach to the Fluorescence Imaging of an Enzyme-Based Biomarker",

abstract = "Here, we report a new pentafluoropropanamido rhodamine fluorescent probe (ACS-HNE) that allows for the selective detection of neutrophil elastase (NE). ACS-HNE displayed high sensitivity, with a low limit of detection (<5.3 nM), and excellent selectivity toward elastase over other relevant biological analytes and enzymes. The comparatively poor solubility and cell permeability of neat ACS-HNE was improved by creating an ACS-HNE-albumin complex; this approach allowed for improvements in the in situ visualization of elastase activity in RAW 264.7 cells relative to ACS-HNE alone. The present study thus serves to demonstrate a simple universal strategy that may be used to overcome cell impermeability and solubility limitations, and to prepare probes suitable for the cellular imaging of enzymatic activity in vitro.",

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AU - Williams, George T.

AU - Gwynne, Lauren

AU - Brewster, James T.

AU - Bull, Steven D.

AU - Jenkins, A. Toby A.

AU - He, Xiao Peng

AU - Schönherr, Holger

AU - Sessler, Jonathan L.

AU - James, Tony D.

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AB - Here, we report a new pentafluoropropanamido rhodamine fluorescent probe (ACS-HNE) that allows for the selective detection of neutrophil elastase (NE). ACS-HNE displayed high sensitivity, with a low limit of detection (<5.3 nM), and excellent selectivity toward elastase over other relevant biological analytes and enzymes. The comparatively poor solubility and cell permeability of neat ACS-HNE was improved by creating an ACS-HNE-albumin complex; this approach allowed for improvements in the in situ visualization of elastase activity in RAW 264.7 cells relative to ACS-HNE alone. The present study thus serves to demonstrate a simple universal strategy that may be used to overcome cell impermeability and solubility limitations, and to prepare probes suitable for the cellular imaging of enzymatic activity in vitro.

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Protein Encapsulation: A Nanocarrier Approach to the Fluorescence Imaging of an Enzyme-Based Biomarker

Abstract

Funding

Keywords

ASJC Scopus subject areas

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