Production and evaluation of the utility of novel phage display-derived peptide ligands to Salmonella spp. for magnetic separation
Article first published online: 15 APR 2013
Journal of Applied Microbiology © 2013 The Society for Applied Microbiology
Journal of Applied Microbiology
Volume 115, Issue 1, pages 271–281, July 2013
How to Cite
Morton, J., Karoonuthaisiri, N., Stewart, L.D., Oplatowska, M., Elliott, C.T. and Grant, I.R. (2013), Production and evaluation of the utility of novel phage display-derived peptide ligands to Salmonella spp. for magnetic separation. Journal of Applied Microbiology, 115: 271–281. doi: 10.1111/jam.12207
- Issue published online: 12 JUN 2013
- Article first published online: 15 APR 2013
- Accepted manuscript online: 1 APR 2013 09:38AM EST
- Manuscript Accepted: 24 MAR 2013
- Manuscript Revised: 22 MAR 2013
- Manuscript Received: 7 JAN 2013
- EU FP7 Incoming Marie Curie
The objectives of this study were to produce Salmonella-specific peptide ligands by phage display biopanning and evaluate their use for magnetic separation (MS).
Methods and Results
Four-phage display biopanning rounds were performed, and the peptides expressed by the two most Salmonella-specific (on the basis of phage-binding ELISA results) phage clones, MSal020401 and MSal020417, were chemically synthesized and coupled to MyOne™ tosylactivated Dynabeads®. Peptide capture capability for whole Salmonella cells from nonenriched broth cultures was quantified by MS + plate counts and MS + Greenlight™ detection and compared to capture capability of anti-Salmonella (antibody-coated) Dynabeads®. MS + Greenlight™ gave a more comprehensive picture of capture capability than MS + plate counts and showed that Peptide MSal020417-coated beads exhibited at least similar, if not better, capture capability to anti-Salmonella Dynabeads® (mean capture values of 36·0 ± 18·2 and 31·2 ± 20·1%, respectively, over Salmonella spp. concentration range 3 × 101–3 × 106 CFU ml−1) with cross-reactivity of ≤1·9% to three other foodborne pathogens: Escherichia coli, Listeria monocytogenes and Campylobacter jejuni.
One of the phage display-derived peptide ligands was demonstrated by MS + Greenlight™ to be a viable antibody alternative for MS of Salmonella spp.
Significance and Impact of the Study
This study demonstrates an antibody-free approach to Salmonella detection and opens substantial possibilities for more rapid tests for this bacterium.