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Biological Activity of Masked Endotoxin
Low endotoxin recovery (LER) is a recently discovered phenomenon describing the inability of limulus amebocyte lysate (LAL)-based assays to detect lipopolysaccharide (LPS) because of a “masking effect” caused by chelators or detergents commonly used in buffer formulations for medical products and re...
Autores principales: | , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2017
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5357793/ https://www.ncbi.nlm.nih.gov/pubmed/28317862 http://dx.doi.org/10.1038/srep44750 |
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author | Schwarz, Harald Gornicec, Jan Neuper, Theresa Parigiani, Maria Alejandra Wallner, Michael Duschl, Albert Horejs-Hoeck, Jutta |
author_facet | Schwarz, Harald Gornicec, Jan Neuper, Theresa Parigiani, Maria Alejandra Wallner, Michael Duschl, Albert Horejs-Hoeck, Jutta |
author_sort | Schwarz, Harald |
collection | PubMed |
description | Low endotoxin recovery (LER) is a recently discovered phenomenon describing the inability of limulus amebocyte lysate (LAL)-based assays to detect lipopolysaccharide (LPS) because of a “masking effect” caused by chelators or detergents commonly used in buffer formulations for medical products and recombinant proteins. This study investigates the masking capacities of different buffer formulations and whether masked endotoxin is biologically active. We show that both naturally occurring endotoxin as well as control standard endotoxin can be affected by LER. Furthermore, whereas masked endotoxin cannot be detected in Factor C based assays, it is still detectable in a cell-based TLR4-NF-κB-luciferase reporter gene assay. Moreover, in primary human monocytes, masked LPS induces the expression of pro-inflammatory cytokines and surface activation markers even at very low concentrations. We therefore conclude that masked LPS is a potent trigger of immune responses, which emphasizes the potential danger of masked LPS, as it may pose a health threat in pharmaceutical products or compromise experimental results. |
format | Online Article Text |
id | pubmed-5357793 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2017 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-53577932017-03-22 Biological Activity of Masked Endotoxin Schwarz, Harald Gornicec, Jan Neuper, Theresa Parigiani, Maria Alejandra Wallner, Michael Duschl, Albert Horejs-Hoeck, Jutta Sci Rep Article Low endotoxin recovery (LER) is a recently discovered phenomenon describing the inability of limulus amebocyte lysate (LAL)-based assays to detect lipopolysaccharide (LPS) because of a “masking effect” caused by chelators or detergents commonly used in buffer formulations for medical products and recombinant proteins. This study investigates the masking capacities of different buffer formulations and whether masked endotoxin is biologically active. We show that both naturally occurring endotoxin as well as control standard endotoxin can be affected by LER. Furthermore, whereas masked endotoxin cannot be detected in Factor C based assays, it is still detectable in a cell-based TLR4-NF-κB-luciferase reporter gene assay. Moreover, in primary human monocytes, masked LPS induces the expression of pro-inflammatory cytokines and surface activation markers even at very low concentrations. We therefore conclude that masked LPS is a potent trigger of immune responses, which emphasizes the potential danger of masked LPS, as it may pose a health threat in pharmaceutical products or compromise experimental results. Nature Publishing Group 2017-03-20 /pmc/articles/PMC5357793/ /pubmed/28317862 http://dx.doi.org/10.1038/srep44750 Text en Copyright © 2017, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Schwarz, Harald Gornicec, Jan Neuper, Theresa Parigiani, Maria Alejandra Wallner, Michael Duschl, Albert Horejs-Hoeck, Jutta Biological Activity of Masked Endotoxin |
title | Biological Activity of Masked Endotoxin |
title_full | Biological Activity of Masked Endotoxin |
title_fullStr | Biological Activity of Masked Endotoxin |
title_full_unstemmed | Biological Activity of Masked Endotoxin |
title_short | Biological Activity of Masked Endotoxin |
title_sort | biological activity of masked endotoxin |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5357793/ https://www.ncbi.nlm.nih.gov/pubmed/28317862 http://dx.doi.org/10.1038/srep44750 |
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