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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Acknowledgments
We like to thank Prof. Dr. Karl-Heinz Wiesmüller (EMCmicrocollections, Tübingen, Germany) for providing us with FSL-1. We like to thank Prof. Dr. Ulrich Zähringer (formerly Forschungszentrum Borstel, Germany) for providing us with E. coli LPS. We like to thank Dimitri Kasakovski for generating the cigarette and smoke extracts. This work was supported by Deutsche Forschungsgemeinschaft (PE 1813/2-1).
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Peters, M., Bonowitz, P., Bufe, A. (2017). A Bioassay for the Determination of Lipopolysaccharides and Lipoproteins. In: Holst, O. (eds) Microbial Toxins. Methods in Molecular Biology, vol 1600. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6958-6_14
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DOI: https://doi.org/10.1007/978-1-4939-6958-6_14
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Publisher Name: Humana Press, New York, NY
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