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On-Particle Rolling Circle Amplification-Based Core-Satellite Magnetic Superstructures for MicroRNA Detection
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Solid State Physics.
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 Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology.
BluSense Diagnostics, Copenhagen, Denmark.
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2018 (English)In: ACS Applied Materials and Interfaces, ISSN 1944-8244, E-ISSN 1944-8252, Vol. 10, no 3, p. 2957-2964Article in journal (Refereed) Published
Abstract [en]

Benefiting from the specially tailored properties of the building blocks as well as of the scaffolds, DNA-assembled core satellite superstructures have gained increasing interest-in drug delivery, imaging, and biosensing. The load of satellites plays,,a vital role in core satellite superstructures, and it determines the signal intensity in response to a biological/physical stimulation/actuation. Herein, for the first time, we utilize on-particle rolling circle amplification (RCA) to prepare rapidly responsive-core satellite magnetic superstructures With a high load of magnetic nanoparticle (MNP) Satellites. Combined with duplex-specific nuclease-assisted target recycling) the proposed magnetic superstructures hold great promise in sensitive and rapid microRNA detection. The long single-stranded DNA produced by RCA serving as the scaffold of the core satellite superstructure can be hydrolyzed by duplex-Specific nuclease in the presence of target microRNA, resulting in a release of MNPs that can be quantified in an optomagnetic sensor. The proposed biosensor has a-simple mix separate measure strategy. For let-7b detection, the proposed biosensor offers a wide linear detection range of approximately 5 orders of magnitude with a detection sensitivity of 1 fM. Moreover, it has the capability to discriminate single-nucleotide mismatches and to detect let-7b in cell extracts and serum, thus showing considerable potential for clinical applications.

Place, publisher, year, edition, pages
2018. Vol. 10, no 3, p. 2957-2964
National Category
Engineering and Technology
Research subject
Engineering Science with specialization in Solid State Physics
Identifiers
URN: urn:nbn:se:uu:diva-345594DOI: 10.1021/acsami.7b16293ISI: 000423496500087PubMedID: 29266917OAI: oai:DiVA.org:uu-345594DiVA, id: diva2:1189359
Funder
Swedish Research Council Formas, 221-2012-444Available from: 2018-03-09 Created: 2018-03-09 Last updated: 2018-03-20Bibliographically approved
In thesis
1. Magnetic Nanoparticle Based Biosensors for Pathogen Detection and Cancer Diagnostics
Open this publication in new window or tab >>Magnetic Nanoparticle Based Biosensors for Pathogen Detection and Cancer Diagnostics
2018 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

This thesis describes several magnetic nanoparticle (MNP)-based biosensing strategies which take advantage of different magnetic sensors, molecular tools and nanotechnologies. Proposed biosensors can be classified into three groups, i.e., immunoassay-based, molecular amplification-based, and nanoparticle assembly-based. The principal motivation is to develop and optimize biosensors for out-of-lab and point-of-care testing.

Immunoassay-based biosensors described in this thesis employ antibodies as the bio-recognition element for the detection of bacteria cells/fragments or proteins. Two typical immunoassay formats, i.e., direct and competitive format, are studied and compared for bacteria detection. In addition, in the protein biomarker detection, MNP chains are formed in the presence of target analytes as well as in the external rotating magnetic field. The high shape/magnetic anisotropy of the chains provides better optomagnetic performance.

Two different molecular amplification methods, i.e., rolling circle amplification (RCA) and loop-mediated isothermal amplification (LAMP), are described under the topic of molecular amplification-based biosensors. In RCA-based biosensors, DNA probe modified MNPs bind to the amplicons after amplification. In LAMP-based biosensors, MNPs are either modified with primers that keep growing during the amplification, or are co-precipitated with the by-product (Mg2P2O7) of the amplification.

The design of the nanoparticle assembly-based biosensors described in this thesis is based on duplex-specific nuclease (DSN)-assisted target recycling and core-satellite magnetic superstructures. In the presence of target microRNA, DSN cuts the DNA scaffold of the core-satellite assembly, releasing MNP satellites that can be quantified by the sensor.

Different kinds of target analytes, i.e., pathogens or cancer biomarkers, are detected at the aiming for rapid, low-cost and user-friendly diagnostics.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2018. p. 55
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1647
Keywords
Magnetic biosensors, magnetic nanoparticles, homogeneous assays, volumetric sensing
National Category
Other Engineering and Technologies not elsewhere specified
Research subject
Engineering Science with specialization in Solid State Physics
Identifiers
urn:nbn:se:uu:diva-346014 (URN)978-91-513-0278-2 (ISBN)
Public defence
2018-05-04, Häggsalen, Ångströmlaboratoriet, Lägerhyddsv. 1, Uppsala, 13:15 (English)
Opponent
Supervisors
Available from: 2018-04-13 Created: 2018-03-13 Last updated: 2018-04-24

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Tian, BoQiu, ZhenMa, JingSvedlindh, PeterStrömberg, Mattias

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