A bioassay for the determination of lipopolysaccharides and lipoproteins

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.