Production and evaluation of the utility of novel phage display-derived peptide ligands to Salmonella spp. for magnetic separation

J Morton, N Karoonuthaisiri, L D Stewart, M Oplatowska, C T Elliott, I R Grant

Research output: Contribution to journalArticlepeer-review

19 Citations (Scopus)

Abstract

Aims: The objectives of this study were to produce Salmonella-specific peptideligands by phage display biopanning and evaluate their use for magneticseparation (MS).
Methods and Results: Four-phage display biopanning rounds were performed,and the peptides expressed by the two most Salmonella-specific (on the basisof phage-binding ELISA results) phage clones, MSal020401 and MSal020417,were chemically synthesized and coupled to MyOneTMtosylactivatedDynabeadsâ. Peptide capture capability for whole Salmonella cells fromnonenriched broth cultures was quantified by MS + plate counts andMS + GreenlightTMdetection and compared to capture capability of anti-Salmonella (antibody-coated) Dynabeadsâ.MS+ GreenlightTMgave a morecomprehensive picture of capture capability than MS + plate counts andshowed that Peptide MSal020417-coated beads exhibited at least similar, if notbetter, capture capability to anti-Salmonella Dynabeadsâ(mean capture valuesof 360  182 and 312  201%, respectively, over Salmonella spp.concentration range 3 9 101–3 9 106CFU ml1) with cross-reactivity of 19% to three other foodborne pathogens: Escherichia coli, Listeriamonocytogenes and Campylobacter jejuni.
Conclusions: One of the phage display-derived peptide ligands wasdemonstrated by MS + GreenlightTMto be a viable antibody alternative for MSof Salmonella spp.
Significance and Impact of the Study: This study demonstrates an antibody-free approach to Salmonella detection and opens substantial possibilities formore rapid tests for this bacterium.
Original languageEnglish
Pages (from-to)271--281
Number of pages11
JournalJournal of applied microbiology
Volume115
Issue number1
Early online date15 Apr 2013
DOIs
Publication statusPublished - 12 Jun 2013

Keywords

  • Salmonella
  • bacteriophages
  • peptides
  • detection
  • magnetic separation

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Biotechnology

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