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The objective of this study was to produce phage display-derived binders with the ability to distinguish   Listeria monocytogenes   from otherListeria    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 otherListeria    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 otherListeria    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.

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