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MicroRNA detection based on duplex-specific nuclease-assisted target recycling and gold nanoparticle/graphene oxide nanocomposite-mediated electrocatalytic amplification
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Applied Materials Sciences.ORCID iD: 0000-0001-7591-2969
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Physics.ORCID iD: 0000-0002-7892-5260
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Polymer Chemistry.ORCID iD: 0000-0002-7256-0758
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Polymer Chemistry.
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2019 (English)In: Biosensors & bioelectronics, ISSN 0956-5663, E-ISSN 1873-4235, Vol. 127, p. 188-193Article in journal (Refereed) Published
Abstract [en]

DNA technology based bio-responsive nanomaterials have been widely studied as promising tools for biomedical applications. Gold nanoparticles (AuNPs) and graphene oxide (GO) sheets are representative zero- and two-dimensional nanomaterials that have long been combined with DNA technology for point-of-care diagnostics. Herein, a cascade amplification system based on duplex-specific nuclease (DSN)-assisted target recycling and electrocatalytic water-splitting is demonstrated for the detection of microRNA. Target microRNAs can form DNA: RNA heteroduplexes with DNA probes on the surface of AuNPs, which can be hydrolyzed by DSN. MicroRNAs are preserved during the reaction and released into the suspension for the digestion of multiple DNA probes. After the DSN-based reaction, AuNPs are collected and mixed with GO to form AuNP/GO nanocomposite on an electrode for the following electrocatalytic amplification. The utilization of AuNP/GO nanocomposite offers large surface area, exceptional affinity to water molecules, and facilitated mass diffusion for the water-splitting reaction. For let-7b detection, the proposed biosensor achieved a limit detection of 1.5 fM in 80 min with a linear detection range of approximately four orders of magnitude. Moreover, it has the capability of discriminating non-target microRNAs containing even single-nucleotide mismatches, thus holding considerable potential for clinical diagnostics.

Place, publisher, year, edition, pages
2019. Vol. 127, p. 188-193
Keywords [en]
Gold nanoparticles, Graphene oxide, MicroRNA detection, Electrocatalytic amplification, Duplex-specific nuclease
National Category
Analytical Chemistry Physical Chemistry
Identifiers
URN: urn:nbn:se:uu:diva-377203DOI: 10.1016/j.bios.2018.12.027ISI: 000457508800026PubMedID: 30611105OAI: oai:DiVA.org:uu-377203DiVA, id: diva2:1291620
Funder
Swedish Research Council, 2016-05259Knut and Alice Wallenberg FoundationEU, Horizon 2020, 713683Available from: 2019-02-25 Created: 2019-02-25 Last updated: 2019-04-24Bibliographically approved
In thesis
1. Biosensing platforms using graphene based bioreactive nanostructures with various dimensions
Open this publication in new window or tab >>Biosensing platforms using graphene based bioreactive nanostructures with various dimensions
2019 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Nanomaterials have brought new aspects and improvements to the biosensing field due to their unique physical and chemical properties that are not shown in the bulk state. This thesis focuses on concepting, developing and testing of biosensors where nanomaterials including graphene gold nanoparticles (AuNPs) and magnetic nanoparticles (MNPs) constitute the biosensors. The motivation is to improve the properties of biosensors for protein and nucleic acids by using the nanomaterials’ high surface volume ratio, their unique electrical properties, their good stability and biocompatibility.

The synthesis of well controlled hybrid materials was essential to obtain well performing nucleic acids sensors, whereas a protein sensor contained mainly graphene and organic molecules. The nanomechanical measurements were applied on pyrene-maltose functionalized graphene surfaces after incubating them with the protein. When the Concanavalin A was captured by the pyrene-maltose, the adhesion force of biosensor surface increased significantly. This detection principle was employed to quantify the Concanavalin A attachment to the surface sensitively.

In the development of the eletrocatalytic microRNA sensor, AuNPs were packaged into graphene oxide (GO) sheets to form three-dimensional network structures that both guide the electrical current and increase the surface area of the electrodes. Prior to the assembly of these GO-AuNPs hybrid materials, a duplex-specific nuclease-assisted target recycling reaction was employed for opening the surface of the DNA functionalized AuNPs. The electrocatalytical water splitting activity increased with the fraction of the AuNP surface and thus with the activity of the nuclease-assisted target recycling reaction.

Owing to the high shape anisotropy of graphene, a two-dimensional optomagnetic label GO-MNP nanohybrid was investigated for DNA detection. The DNA coils that were generated through rolling circle amplification absorbed on GO-MNP nanohybrid, leading to a hydrodynamic size increase or aggregation of the proposed nanolabels that can be detected by an optomagnetic sensor. An MNP assembly-based microRNA biosensing strategy is also included in the thesis. DNA scaffolds of the MNP assemblies contain DNAzyme substrate and thus can form cleavage catalytic structures in the presence of microRNA, leading to the disintegration of assemblies. The proposed nanomaterials based biosensing platforms show great potential in the clinical and biomedical applications.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2019. p. 55
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1779
Keywords
Graphene, nanoparticles, nanostructure, biosensors
National Category
Analytical Chemistry Engineering and Technology
Identifiers
urn:nbn:se:uu:diva-378561 (URN)978-91-513-0587-5 (ISBN)
Public defence
2019-04-25, Room 2005, Ångströmlaboratoriet, Lägerhyddsvägen 1, Uppsala, 13:15 (English)
Opponent
Supervisors
Available from: 2019-04-01 Created: 2019-03-06 Last updated: 2019-05-07

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Han, YuanyuanQiu, ZhenNawale, Ganesh N.Varghese, Oommen P.Hilborn, JönsTian, BoLeifer, Klaus

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