Journal
ANALYTICAL CHEMISTRY
Volume 90, Issue 22, Pages 13600-13606Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.8b03809
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Funding
- National Natural Science Foundation of China [21575158, 21475148, 41876108]
- Strategic Priority Research Program of the Chinese Academy of Sciences [XDA11020702]
- Taishan Scholar Program of Shandong Province [TS20081159]
- National Key Research and Development Program of China [2016YFC1400700]
- Key Research and Development Program of Yantai [2018ZHGY053]
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Peptide-based sandwich assays are promising tools in molecular detection, but may be restricted by the availability of pairs of affinity peptides. Herein, a new potentiometric sandwich assay for bacteria based on peptide pairs derived from an antimicrobial peptide (AMP) ligand is demonstrated. As a model, the original AMP with a welldefined structure for Listeria monocytogenes (LM) can be split into two fragments to serve as the peptide pairs for the sandwich assay. The recognition and binding of the short peptide pairs to the target can be verified by circular dichroism, flow cytometry, fluorometry, and optical microscopy. The potentiometric magnetic bead-based sandwich assay is designed by using horseradish peroxidase as a label. The enzyme can catalyze the oxidation of 3,3',5,5'-tetramethylbenzidine with H2O2 to induce a potential change on a polymeric membrane ion selective electrode. Under optimal conditions, the concentration of LM can be determined potentiometrically in a linear range of 1.0 X 10(2) to 1.0 x 10(6) CFU mL(-1) with a detection limit of 10 CFU mL(-1) (3 sigma). The proposed sensing strategy expands the applications of peptides in the field of bioassays.
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