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  • Title: Supersandwich-type electrochemiluminescenct aptasensor based on Ru(phen)3(2+) functionalized hollow gold nanoparticles as signal-amplifying tags.
    Author: Gui G, Zhuo Y, Chai YQ, Liao N, Zhao M, Han J, Zhu Q, Yuan R, Xiang Y.
    Journal: Biosens Bioelectron; 2013 Sep 15; 47():524-9. PubMed ID: 23643946.
    Abstract:
    An electrochemiluminescent (ECL) aptasensor was fabricated and used for the amplified detection of thrombin (TB) based on DNA supersandwich structure. Herein, hollow gold nanospheres (HGNPs) were firstly employed as effective tag-carriers for the immobilization of detection aptamer (TBA 2) to form the HGNPs labeled TBA 2 (HGNPs-TBA 2). Subsequently, streptavidin (SA) was used to block the non-specific binding sites of HGNPs-TBA 2 as well as to supply binding sites, which could further introduce numerous initiator DNA strands (bio-S1) via biotin-streptavidin specific interaction. Next, bio-S1 could in situ trigger hybridization chain reaction (HCR) to create a long nicked double helices analogous (dsDNA) in the present of ssDNA S2 and ssDNA S3 (S3 is partially complementary to the S2) to obtain the DNA supersandwich structure. Furthermore, Ru(phen)3(2+), a well-known ECL luminophore, could be intercalated into the grooves of dsDNA (Ru-dsDNA) to form the Ru-dsDNA-SA-HGNPs-TBA 2 bioconjugate. As a result, the target of TB was sandwiched between Ru-dsDNA-SA-HGNPs-TBA 2 and TBA 1. In this strategy, numerous Ru(phen)3(2+) could be immobilized on the electrode based on the supersandwich structure, resulting in an increased ECL signal output. A supersandwich ECL assay for TB detection was developed with excellent sensitivity of a large concentration variation from 5fM to 50pM and a detection limit of 1.6fM.
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