Journal
ANALYTICAL CHEMISTRY
Volume 89, Issue 11, Pages 5901-5909Publisher
AMER CHEMICAL SOC
DOI: 10.1021/acs.analchem.7b00261
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Funding
- National Natural Foundation of China [21405077, 21227009, 21475060]
- Natural Science Foundation of Jiangsu Province [BK20140591]
- Special Fund for Agro-scientific research in the Public interest [201403071]
- National Science Fund for Creative Research Groups [21121091]
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Here we report a novel type of bivalent aptasensor based on silver-enhanced fluorescence polarization (FP) for detection of lactoferrin (Lac) in milk powder with high sensitivity and specificity. The novel two split aptamers were obtained from the aptamer reported in our previous SELEX (systematic evolution of ligands by exponential enrichment) selection, and their minimal structural units were optimized on the basis of their affinity and specificity. Also, dual binding sites of split aptamers were verified. The bivalent aptamers were modified to be linked with signal molecule fluorescein isothiocyanate (FITC) and enhancer silver decahedral nanoparticles (Ag(10)NPs). The split aptamers could bind to different sites of Lac and assemble into a split-aptamers-target complex, narrowing the distance between Ag10NPs and FITC dye. As a result, Ag10NPs could produce a mass augmented and metal-enhanced fluorescence (MEF) effect. In general, ternary amplification based on Ag10NPs, split aptamers, and the MEF effect all contributed to the significant increase of FP values. It was proved that the sensitivity of this assay was about 3 orders of magnitude over traditional aptamer-based homogeneous assays with a detection limit of 1.25 pM. Furthermore, this design was examined by actual milk powder with rapid and high-throughout detection.
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