Terminal Protection of Small-Molecule-Linked DNA for Sensitive Electrochemical Detection of Protein Binding via Selective Carbon Nanotube Assembly
journal contributionposted on 2009-09-02, 00:00 authored by Zhan Wu, Zhen Zhen, Jian-Hui Jiang, Guo-Li Shen, Ru-Qin Yu
Small-molecule-linked DNA has emerged as a versatile tool for the interaction assay between small organic molecules and their protein receptors. We report herein the proof-of-principle of a terminal protection assay of small-molecule-linked DNA. This assay is based on our new finding that single-stranded DNA (ssDNA) terminally tethered to a small molecule is protected from the degradation by exonuclease I (Exo I) when the small molecule moiety is bound to its protein target. This finding translates the binding of small molecules to proteins into the presence of a specific DNA sequence, which enables us to probe the interaction between small organic molecules and their protein targets using various DNA sequence amplification and detection technologies. On the basis of selective assembly of single-walled carbon nanotubes (SWNTs) with surface-tethered small-molecule-linked ssDNA not protected by protein binding, a novel electrochemical strategy for terminal protection assay has been developed. Through detecting the redox signal mediated by SWNT assembly on a 16-mercaptohexadecanoic acid-blocked electrode, this strategy is able to ensure substantial signal amplification and a low background current. This strategy is demonstrated for quantitative analysis of the interaction of folate with a tumor biomarker of folate receptor (FR), and a detection limit of 3 pM FR is readily achieved with desirable specificity and sensitivity, indicating that the terminal protection assay can offer a promising platform for small molecule−protein interaction studies.
SWNT assemblySensitive Electrochemical Detectionfolate receptorprotein receptorsinteraction assaydetection technologies3 pM FRprotein bindingsignal amplificationmolecule moietyterminal protection assayDNA sequence amplificationDNA sequenceprotein targetdetection limitnovel electrochemical strategyredox signalprotein targetsterminal Protectiontumor biomarkerProtein Binding