Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules

Abstract Every year pharmaceutical companies use significant resources to mitigate aggregation of pharmaceutical drug products. Specifically, peptides and proteins that have been denatured or degraded can lead to adverse patient reactions such as undesired immune responses. Current methods to detect...

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Main Authors: Monique J. Farrell, Robert J. Reaume, Aswini K. Pradhan
Format: Article
Language:English
Published: Nature Publishing Group 2017-06-01
Series:Scientific Reports
Online Access:https://doi.org/10.1038/s41598-017-02899-3
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spelling doaj-a6c447a3d1d644eabf20065780207db12020-12-08T01:27:01ZengNature Publishing GroupScientific Reports2045-23222017-06-017111110.1038/s41598-017-02899-3Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed BiomoleculesMonique J. Farrell0Robert J. Reaume1Aswini K. Pradhan2Center for Materials Research, Norfolk State UniversityCenter for Materials Research, Norfolk State UniversityCenter for Materials Research, Norfolk State UniversityAbstract Every year pharmaceutical companies use significant resources to mitigate aggregation of pharmaceutical drug products. Specifically, peptides and proteins that have been denatured or degraded can lead to adverse patient reactions such as undesired immune responses. Current methods to detect aggregation of biological molecules are limited to costly and time consuming processes such as high pressure liquid chromatography, ultrahigh pressure liquid chromatography and SDS-PAGE gels. Aggregation of pharmaceutical drug products can occur during manufacturing, processing, packaging, shipment and storage. Therefore, a facile in solution detection method was evaluated to visually detect denatured glutathione peptides, utilizing gold nanoparticle aggregation via 3-Aminopropyltreithoxysilane. Glutathione was denatured using a 70 °C water bath to create an accelerated heat stressed environment. The peptide, gold nanoparticle and aminosilane solution was then characterized via, UV-Vis spectroscopy, FTIR spectroscopy, dynamic light scattering and scanning electron microscopy. Captured images and resulting absorbance spectra of the gold nanoparticle, glutathione, and aminosilane complex demonstrated visual color changes detectable with the human eye as a function of the denaturation time. This work serves as an extended proof of concept for fast in solution detection methods for glutathione peptides that have experienced heat stress.https://doi.org/10.1038/s41598-017-02899-3
collection DOAJ
language English
format Article
sources DOAJ
author Monique J. Farrell
Robert J. Reaume
Aswini K. Pradhan
spellingShingle Monique J. Farrell
Robert J. Reaume
Aswini K. Pradhan
Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules
Scientific Reports
author_facet Monique J. Farrell
Robert J. Reaume
Aswini K. Pradhan
author_sort Monique J. Farrell
title Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules
title_short Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules
title_full Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules
title_fullStr Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules
title_full_unstemmed Visual Detection of Denatured Glutathione Peptides: A Facile Method to Visibly Detect Heat Stressed Biomolecules
title_sort visual detection of denatured glutathione peptides: a facile method to visibly detect heat stressed biomolecules
publisher Nature Publishing Group
series Scientific Reports
issn 2045-2322
publishDate 2017-06-01
description Abstract Every year pharmaceutical companies use significant resources to mitigate aggregation of pharmaceutical drug products. Specifically, peptides and proteins that have been denatured or degraded can lead to adverse patient reactions such as undesired immune responses. Current methods to detect aggregation of biological molecules are limited to costly and time consuming processes such as high pressure liquid chromatography, ultrahigh pressure liquid chromatography and SDS-PAGE gels. Aggregation of pharmaceutical drug products can occur during manufacturing, processing, packaging, shipment and storage. Therefore, a facile in solution detection method was evaluated to visually detect denatured glutathione peptides, utilizing gold nanoparticle aggregation via 3-Aminopropyltreithoxysilane. Glutathione was denatured using a 70 °C water bath to create an accelerated heat stressed environment. The peptide, gold nanoparticle and aminosilane solution was then characterized via, UV-Vis spectroscopy, FTIR spectroscopy, dynamic light scattering and scanning electron microscopy. Captured images and resulting absorbance spectra of the gold nanoparticle, glutathione, and aminosilane complex demonstrated visual color changes detectable with the human eye as a function of the denaturation time. This work serves as an extended proof of concept for fast in solution detection methods for glutathione peptides that have experienced heat stress.
url https://doi.org/10.1038/s41598-017-02899-3
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