Submitted to: UJNR Food & Agricultural Panel Proceedings
Publication Type: Proceedings
Publication Acceptance Date: 10/20/2004
Publication Date: 12/14/2004
Citation: Paoli, G., Brewster, J.D., Kleina, L.G. 2004. Development of biosensors from phage displayed antibodies. UJNR Food & Agricultural Panel Proceedings. pp. 359-364.
Technical Abstract: Phage display is a useful technique for the isolation of antibody fragments with desired specificities. We have selected and screened a phage displayed single-chain antibody (scFv) library to isolate an antibody fragment that detects several strains of L. monocytogenes, and does not cross-react with any of the other five species of Listeria. The effectiveness of this antibody fragment for the detection of L.monocytogenes was examined by ELISA using L. monocytogenes grown in commonly used enrichment media and at a variety of temperatures. The antibody bound to a polypeptide antigen with a molecular mass of 100 kD that was both on the surface of L. monocytogenes cells and exported out of the cells. The antigen was identified by MALDI-TOF mass spectrometry. We are making a set of genetic constructs to fuse antibody fragments to appropriate protein tags and reporters in order to develop immuno-biosensors for the detection of food-borne pathogens.