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

Abstract

The objective of this study was to produce phage display-derived binders with the ability to distinguish Listeria monocytogenes from other Listeria spp., which may have potential utility to enhance detection of Listeria monocytogenes. To obtain binders with the desired binding specificity a series of surface and solution phage-display biopannings were performed. Initially, three rounds of surface biopanning against gamma-irradiated L. monocytogenes serovar 4b cells were performed followed by an additional surface biopanning round against L. monocytogenes 4b which included prior subtraction biopanning against gamma-irradiated L. innocua cells. In an attempt to further enhance binder specificity for L. monocytogenes 4b two rounds of solution biopanning were performed, both rounds included initial subtraction solution biopanning against L. innocua. Subsequent evaluations were performed on the phage clones by phage binding ELISA. All phage clones tested from the second round of solution biopanning had higher specificity for L. monocytogenes 4b than for L. innocua and three other foodborne pathogens ( Salmonella spp., Escherichia coli and Campylobacter jejuni). Further evaluation with five other Listeria spp. revealed that one phage clone in particular, expressing peptide GRIADLPPLKPN, was highly specific for L. monocytogenes with at least 43-fold more binding capability to L. monocytogenes 4b than to any other Listeria sp. This proof-of-principle study demonstrates how a combination of surface, solution and subtractive biopanning was used to maximise binder specificity. L. monocytogenes-specific binders were obtained which could have potential application in novel detection tests for L. monocytogenes, benefiting both the food and medical industries.

Isolation of phage-display library-derived scFv antibody specific toListeria monocytogenesby a novel immobilized method

“…; Morton et al . ; Tu et al . ), the developed antibodies for L. monocytogenes either cross‐reacted with other Listeria species or failed to bind to a wide range of L. monocytogenes strains.…”

“…; Morton et al . ; Tu et al . ); however, this is the first time a phage clone has been reported to recognize not only a wide range of pathogenic L. monocytogenes serotypes but also different strains belonging to the same serotype.…”

“…Despite the many attempts made to isolate L. monocytogenes-specific antibodies (Bhunia et al 1991;Nannapaneni et al 1998;Paoli et al 2004;Carnazza et al 2007;Morton et al 2013;Tu et al 2016), the developed antibodies for L. monocytogenes either cross-reacted with other Listeria species or failed to bind to a wide range of L. monocytogenes strains.…”

“…Therefore, it is important to examine the isolated binders over not only a wide range of serotypes but also different strains belonging to the same serotype. Many attempts have been made to produce L. monocytogenes-specific binders from phage-display libraries (Paoli et al 2004;Carnazza et al 2007;Morton et al 2013;Tu et al 2016); however, this is the first time a phage clone has been reported to recognize not only a wide range of pathogenic L. monocytogenes serotypes but also different strains belonging to the same serotype.…”

Listeria monocytogenes in the Food Processing Environment

“…Differentiation of L. monocytogenes as distinct from other Listeria species has also been seen to be possible through phage typing. As different bacterial species are susceptible to different phages, the use of a specifi c phage which has a known, predictable species target can identify L. monocytogenes (Morton et al 2013 ).…”