Phage display-derived binders able to distinguish Listeria monocytogenes from other Listeria species

• Published in: PloS one Vol. 8; no. 9; p. e74312
• Main Authors: McIvor, Mary Josephine, Karoonuthaisiri, Nitsara, Charlermroj, Ratthaphol, Stewart, Linda D, Elliott, Christopher T, Grant, Irene R
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 10-09-2013; Public Library of Science (PLoS)
• Subjects: Amino Acid Sequence; Animals; Antigens; Antigens, Bacterial - chemistry; Antigens, Bacterial - metabolism; B cells; Bacteria; Bacterial Typing Techniques - methods; Binders; Binding; Biosensors; Campylobacter; Campylobacter jejuni - chemistry; Deoxyribonucleic acid; DNA; E coli; Enzyme-linked immunosorbent assay; Enzyme-Linked Immunosorbent Assay - methods; Epidemics; Escherichia coli; Escherichia coli - chemistry; Food industry; Food processing industry; Food safety; Food supply; Foodborne pathogens; Gamma Rays; Genetic engineering; Immunoglobulins; Immunology; Laboratories; Ligands; Listeria; Listeria - chemistry; Listeria - radiation effects; Listeria monocytogenes; Listeria monocytogenes - chemistry; Listeria monocytogenes - isolation & purification; Listeria monocytogenes - radiation effects; Molecular Sequence Data; Pathogens; Peptide Library; Peptides; Phage display; Phages; Protein Binding; Salmonella; Salmonella - chemistry; Species Specificity; Subtraction; Zoonoses; United Kingdom > UK; Northern Ireland
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0074312

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.