A LISTERIA MONOCYTOGENES‐SPECIFIC PHAGE‐DISPLAYED ANTIBODY FRAGMENT RECOGNIZES A CELL SURFACE PROTEIN WHOSE EXPRESSION IS REGULATED BY PHYSIOLOGICAL CONDITIONS¹

Abstract

We recently selected a phage-displayed single-chain antibody that detects several strains of Listeria monocytogenes and does not cross-react with any of the other five species of Listeria. The efficacy of a phage-displayed anti-L. monocytogenes single-chain antibody as a detection reagent was examined by enzyme-linked immunosorbent assay using L. monocytogenes grown at different temperatures and in a variety of media commonly used for the isolation of L. monocytogenes from food. The best results were observed when cells were grown in supplemented Fraser enrichment broth. The antigen detected by the phage-displayed antibody was present on the surface of the cells grown between 20 and 42C, but was not present on the cell surface when cells were grown at or below 15C. As determined by Western blot analysis, the antibody bound to a protein with apparent molecular mass of approximately 90 kD. Identification of the protein antigen would allow a more rational approach to the development of an antibody-based method for the specific detection of L. monocytogenes. PRACTICAL APPLICATIONSAntibody phage display is an alternative approach to the selection of immunoreagents for the detection of food-borne pathogens. Recently, a phage-1 Mention of brand or firm names does not constitute an endorsement by the U.S. Department of Agriculture over others of a similar nature not mentioned. 2 Corresponding 77 displayed antibody that detected several strains of Listeria monocytogenes, and did not cross-react with any of the other five species of Listeria, was selected from an antibody phage display library. In this work, the efficacy of this phage-displayed antibody for the detection of L. monocytogenes was examined using cells grown under a variety of conditions. In addition, the target that is detected by the phage-displayed antibody was characterized. Characterization of the L. monocytogenes-specific phage-displayed antibody fragment will allow a more rational approach toward the development of methods for the rapid and specific detection of L. monocytogenes from food.

Fructose 1,6-Bisphosphate Aldolase, a Novel Immunogenic Surface Protein on Listeria Species

“…Antibodies have been widely used in immunological tests for specific detection and identification of pathogens [ 19 – 22 ]. The availability of monoclonal antibodies (mAbs) against bacterial surface antigens not only allows the development of detection assays but also provide a powerful tool for the study of bacterial protein structures and functions [ 23 – 27 ]. Since mAbs recognize exclusive epitopes in the antigen, they can be used to identify new proteins that would be important in the bacterial pathogenesis, survival, or adaptation to the environment [ 28 ].…”

SPR biosensor for the detection of L. monocytogenes using phage-displayed antibody

“…The phage probe used in this study, Lm P4:A8, was selected from a naive scFv phage display library for its specific interaction with L. monocytogenes (Paoli et al, 2004). The scFv expressed on the surface of phage Lm P4:A8 interacts with ActA (Paoli and Brewster, 2007), an L. monocytogenes protein virulence factor that is expressed on the bacterial cell surface and functions in host cell-to-cell movement via actin polymerization.…”

Phage Display-Derived Binders Able to Distinguish Listeria monocytogenes from Other Listeria Species

“…This technique has been used previously in an attempt to generate alternative binders to L. monocytogenes ; Paoli et al [5,23] and Nanduri et al [24] produced phage display-derived antibody fragments to L. monocytogenes , while Carnazza et al [25] described the production of phage display-derived peptide binders to L.…”