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Inhibitory Effect of Target Binding on Hairpin Aptamer Sticky-End Pairing-Induced Gold Nanoparticle Assembly for Light-up Colorimetric Protein Assay
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文摘
Gold nanoparticles (GNPs) possessing strong distance-dependent optical properties and high extinction coefficients have emerged as important colorimetric materials. Almost all colorimetric studies are based on two working mechanisms: sandwich cross-linking and non-cross-linking systems. In the present study, a new working mechanism, hairpin sticky-end pairing-induced GNP assembly, is introduced based on the discovery of unique aggregation behavior of aptamer-functionalized GNPs. The salt-induced aggregation of oligonucleotide probe-modified GNPs can readily occur due to the sticky-end pairing effect while addition of target molecules favors the formation of the hairpin structure of probe sequences and substantially inhibits the nanoparticle assembly. Along this line, we developed a proof-of-concept colorimetric homogeneous assay using immunoglobulin E (IgE) as an analyte model via transforming a commonly designed “light-down” colorimetric biosensor into a “light-up” one. From the point of view of both conformational transition of aptamer and steric bulk, oligonucleotide−GNPs display an additional stability upon binding to target molecules. The assay showed an extremely high sensitivity from both naked eye observations and absorbance measurements. Compared with almost all existing IgE sensing strategies, the proposed colorimetric system possesses a substantially improved analytical performance. Investigating the assembly behavior of hairpin aptamer-modified GNPs could offer new insight into the dependence of the GNP properties on the structure switching and open a new way to design signaling probes and develop colorimetric assay schemes.

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