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Publications (3)16.4 Total impact

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    ABSTRACT: Horseradish peroxidase mimicking DNAzyme (HRP-DNAzyme) attracts growing interest as an amplifying label for biorecognition and biosensing events, especially for DNA detection. However, in the traditional designs, one target molecule can only generate one HRP-DNAzyme, which limits the signal enhancement and thus its sensitivity. In this article, we propose an amplified and label-free colorimetric DNA detection strategy based on nicking endonuclease (NEase)-assisted activation of HRP-DNAzymes (NEAA-DNAzymes). This new strategy relies on the hairpin-DNAzyme probe and NEase-assisted target recycling. In the hairpin-DNAzyme probe, the HRP-DNAzyme sequence is protected in a "caged" inactive structure, whereas the loop region includes the target complementary sequence. Upon hybridization with target, the beacon is opened, resulting in the activation of the HRP-DNAzyme. Meanwhile, upon formation of the duplex, the NEase recognizes a specific nucleotide sequence and cleaves the hairpin-DNAzyme probe into two fragments. After nicking, the fragments of the hairpin-DNAzyme probe spontaneously dissociate from the target DNA. Amplification is accomplished by another hairpin-DNAzyme probe hybridizing to the released intact target to continue the strand-scission cycle, which results in activation of numerous DNAzymes. The activated HRP-DNAzymes generate colorimetric or chemiluminescence readout signals, thus providing the amplified detection of DNA. The detection limit of the colorimetric method is 10 pmol/L, which are three orders of magnitude lower than that without NEase. In addition, the detection limit of the chemiluminescence method is 0.2 pmol/L. Meanwhile, this strategy also exhibits high discrimination ability even against single-base mismatch.
    Talanta 07/2011; 85(1):91-6. · 3.50 Impact Factor
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    ABSTRACT: In this article, we report the first graphene oxide (GO)-based platform to detect protease activity in a homogeneous real-time format. In designing such GO-based biosensing platform, we put a protease substrate peptide as the linker between the energy transfer donor (QDs) and the energy transfer acceptor (GO) to fabricate the GO-peptide-QDs nanoprobes. In the nanoprobes, the photoluminescence (PL) of donor QDs was strongly quenched due to the presence of GO in close proximity. The protease activity caused modulation in the efficiency of the energy transfer between the acceptor and donor, thus enabling the protease assay. The proposed GO-based platform is easy to assemble and has little background interference, yet still give superior sensitivity and rapid response. Furthermore, this GO-QDs architecture can serve as a universal platform by simply changing the types of peptide sequences for the different proteases. In this work, GO-based platform has been successfully applied in the sensitive detection of matrix metalloproteinase (MMP) and thrombin activity. Meanwhile, we also utilized this platform to monitor the protease inhibitor. The proposed GO-based platform is anticipated to find applications in the diagnosis of protease-related diseases and screening of potential drugs with high sensitivity in a high-throughput way.
    Biosensors & Bioelectronics 03/2011; · 6.45 Impact Factor
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    ABSTRACT: In this work, we describe a simple, inexpensive and fast method for the generation of molecularly imprinted polymer (MIP) film on quartz crystal microbalance (QCM) crystals using mussel-inspired polymer. Commonly known as a neurotransmitter, dopamine is also a small-molecule mimic of the adhesive proteins of mussels. Polymerization of dopamine in the presence of template molecule (1,3,5-pentanetricarboxylic acid, an analogue of domoic acid, in this case) could produce an adherent molecularly imprinted polydopamine film coating on QCM crystals. Advantages, such as high hydrophilicity, high biocompatibility and controllable thickness, make this molecularly imprinted polydopamine film an attractive recognition element for sensors. Selective rebinding of domoic acid on mussel-inspired molecularly imprinted polymer (m-MIP) coated crystal was observed as a frequency shift quantified by piezoelectric microgravimetry with the QCM system. The decreasing frequency shows a good linear relationship with the concentration of domoic acid. The quantitation limit of domoic acid was 5 ppb with the linear range of 0-100 ppb. The QCM sensor has high selectivity and was able to distinguish domoic acid from its analogous p-phthalic acid and o-phthalic acid owing to the molecular imprinting effect. In addition, the practical analytical performance of the sensor was examined by evaluating the detection of domoic acid in mussel extracts with satisfactory results. It is envisaged that m-MIP could be suitable as recognition element for sensors and the proposed m-MIP QCM sensor could be employed to detect analyte of interest in complex matrices.
    Biosensors & Bioelectronics 10/2010; 26(2):585-9. · 6.45 Impact Factor