Lapitan, LDS and Zhou, D orcid.org/0000-0003-3314-9242 (2020) A simple magnetic nanoparticle-poly-enzyme nanobead sandwich assay for direct, ultrasensitive DNA detection. In: Kumar, CV, (ed.) Nanoarmoring of Enzymes with Carbon Nanotubes and Magnetic Nanoparticles. Methods in Enzymology, 630 . Academic Press , Oxford, UK , pp. 453-480. ISBN 978-0-12-820143-5
Abstract
A simple magnetic nanoparticle (MNP)-poly-enzyme nanobead sandwich assay for direct detection of ultralow levels of unlabeled target-DNA is developed. This approach uses a capture-DNA covalently linked to a dense PEGylated polymer encapsulated MNP and a biotinylated signal-DNA to sandwich the target-DNA. A DNA ligation is then followed to offer high discrimination between the perfect-match and single-base mismatch target-DNAs. Only the presence of a perfect-match target can covalently link the biotinylated signal-DNA onto the MNP surface for subsequent binding to a polymer nanobead tagged with thousands of copies of high-activity neutravidin-horseradish peroxidase (NAV-HRP) for great enzymatic signal amplification. Combining the advantages of the dense MNP surface PEGylation to reduce non-specific adsorption (assay background) and the powerful signal amplification of poly-enzyme nanobead, this assay can directly quantify the target-DNA down to single digit attomolar with a large linear dynamic range of 5 orders of magnitude (from 10− 18 to 10− 13 M).
Metadata
Item Type: | Book Section |
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Authors/Creators: |
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Editors: |
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Keywords: | Magnetic nanoparticle; Enzymatic assay; Poly-enzyme nanobead; Ultra-sensitivity; DNA detection; Sandwich assay; PEGylation |
Dates: |
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Institution: | The University of Leeds |
Academic Units: | The University of Leeds > Faculty of Engineering & Physical Sciences (Leeds) > School of Chemistry (Leeds) > Inorganic Chemistry (Leeds) |
Depositing User: | Symplectic Publications |
Date Deposited: | 04 Nov 2019 11:50 |
Last Modified: | 14 Feb 2020 14:24 |
Status: | Published |
Publisher: | Academic Press |
Series Name: | Methods in Enzymology |
Identification Number: | 10.1016/bs.mie.2019.10.026 |
Open Archives Initiative ID (OAI ID): | oai:eprints.whiterose.ac.uk:153001 |