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71 Dr.-Ing. Christina Kohl Phone +49 711 970-4183 christina.kohl@igb.fraunhofer.de Dr. rer. nat. Anke Burger-Kentischer Phone +49 711 970-4023 anke.burger-kentischer@ igb.fraunhofer.de Contact Literature [1] Akira, S. et al. (2004) Toll-like receptor signalling, Nat Rev Immunol 4: 499–51 [2] Werner-Felmayer, G. et al. (1995) Detection of bacterial pyro- gens on the basis of their effects on gamma interferon-mediated formation of neopterin or nitrite in cultured monocyte cell lines, Clini Diagn Lab Immunol 2(3): 307–313 orgensen, et al Rapid detection of significant bacte- riuria by use of an automated Limulus amoebocyte lysat assay, J Clin Microbiol 16(3): 587–589 [4] Lakhani, S. A. et al. (2003) Toll-like receptor signaling in sep- sis, Curr Opin Pediatr 15(3): 278–82 [5] Burger-Kentischer, A. et al. (2010) New cell-based innate immune receptor assay for the examination of receptor activity, ligand specificity, signalling pathways and the detection of pyro- gens, J Immunol Meth 358: 93–103 ellbasiertes Testsystem ur Identifi ierung und ifferen ier- ung von Keimspektren (2009) DE 10 2006 031 483; EP 2 041 172 Funding We would like to thank the Fraunhofer-Gesellschaft for funding the project “ImmuStick” under the “Discover” program. Fields of application The ImmuStick is suitable for the legally required proof of the presence of pyrogens in biological products and medical de- vices directive pplication in the classification of sepsis pathogens on patients directly is also conceivable. Since some PPRs also recogni e specific allergens, the Immu tick also offers potential for the detection of allergens in cosmetics, medical technology, the pharmaceutical sector and the food industry. The ImmuStick is the result of further development of an in vitro immunoassay into a cost-effective point-of-use test that can even be used by lay persons and which can also be used as on-site system without the need for complex equipment. The use of receptors of the innate human immune system al- lows all pyrogen classes to be detected. Application-specific further development We have demonstrated the basic feasibility of TLR4 for the detection of LPS. The test system has the potential for modu- lar expansion with additional receptors of the innate immune system (TRL, NOD-like receptors, C-type lectins), in order to specifically adapt the pyrogen spectrum uipped with dif- ferent PRRs, the ImmuStick can detect the complete range of PAMPs quickly, easily and directly on-site. 1 Sample application and result window, biosensor with immobilized TLR4 receptors. 2 Competitive displacement of a weaker binding ligand (red) by LPS (green). 3 Quantitative analysis of the displacement study (see 2). 4 Detection of a sample solution containing pyrogens via test lines in the result window. 0 fluorescenceintensityina.u. red fluorescence (TAMRA) green fluorescence (Alexa Fluor488) 70,000 30,000 20,000 10,000 1. incubation with TAMRA peptide 2. Incubation with LPS Alexa Fluor488 40,000 50,000 60,000 43 Phone +49711970-4183 Phone +49711970-4023 ung von Keimspektren (2009) DE 102006031483; EP 2041172