Chapter title |
A Bioassay for the Determination of Lipopolysaccharides and Lipoproteins
|
---|---|
Chapter number | 14 |
Book title |
Microbial Toxins
|
Published in |
Methods in molecular biology, January 2017
|
DOI | 10.1007/978-1-4939-6958-6_14 |
Pubmed ID | |
Book ISBNs |
978-1-4939-6956-2, 978-1-4939-6958-6
|
Authors |
Marcus Peters, Petra Bonowitz, Albrecht Bufe |
Editors |
Otto Holst |
Abstract |
The availability of convenient assays for the detection and quantification of pathogen-associated molecular patterns (PAMPs) is limited. In the case of lipopolysaccharide (LPS) the so-called LAL (limulus amebocyte lysate) test is available, an assay that is performed with the lysate of the blood of the horse shoe crab. Although a sensitive and convenient assay, it lacks specificity, since it is affected by other endotoxins like, for instance, fungal cell walls as well. Here, we describe a bioassay that can be used to detect and quantitate PAMPs in environmental samples. More specific we demonstrate the usage of TLR2 and TLR4/CD14/MD2 transfected Hek293 cells to quantitatively determine bacterial lipoproteins and LPS, respectively. We show the usefulness of these assays to measure LPS in tobacco before and after combustion. |
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