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Designed protein binders in combination with nanocrystalline diamond for use in high-sensitivity biosensors
Uppsala University, Disciplinary Domain of Medicine and Pharmacy, Faculty of Medicine, Department of Immunology, Genetics and Pathology, Clinical Immunology.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Microsystems Technology.
Uppsala University, Disciplinary Domain of Science and Technology, Technology, Department of Engineering Sciences, Microsystems Technology.
Uppsala University, Disciplinary Domain of Science and Technology, Chemistry, Department of Chemistry - Ångström, Inorganic Chemistry.
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2012 (English)In: Analytical and Bioanalytical Chemistry, ISSN 1618-2642, E-ISSN 1618-2650, Vol. 404, no 6-7, 1643-1651 p.Article in journal (Refereed) Published
Abstract [en]

A platform for diagnostic applications showing signal-to-noise ratios that by far surpass those of traditional bioanalytical test formats has been developed. It combines the properties of modified nanocrystalline diamond (NCD) surfaces and those of polyethylene oxide and polypropylene oxide based block copolymers for surface passivation and binder conjugation with a new class of synthetic binders for proteins. The NCD surfaces were fluorine-, hydrogen-, or oxygen-terminated prior to further biofunctionalization and the surface composition was characterized by X-ray photoelectron spectroscopy. In a proof of principle demonstration targeting the C-reactive protein, an ELISA carried out using an F-terminated diamond surface showed a signal-to-noise ratio of 3,900 which compares well to the signal-to-noise of 89 obtained in an antibody-based ELISA on a polystyrene microtiter plate, a standard test format used in most life science laboratories today. The increase in signal-to-noise ratio is to a large extent the result of extremely efficient passivation of the diamond surface. The results suggest that significant improvements can be obtained in standardized test formats using new materials in combination with new types of chemical coatings and receptor molecules.

Place, publisher, year, edition, pages
2012. Vol. 404, no 6-7, 1643-1651 p.
Keyword [en]
Protein binders, Nanocrystalline diamond, CRP, Biosensor, Surface characterization
National Category
Natural Sciences Engineering and Technology
Research subject
Engineering Science with specialization in Microsystems Technology
Identifiers
URN: urn:nbn:se:uu:diva-184455DOI: 10.1007/s00216-012-6245-7ISI: 000309348400004OAI: oai:DiVA.org:uu-184455DiVA: diva2:566723
Available from: 2012-11-09 Created: 2012-11-07 Last updated: 2017-12-07Bibliographically approved
In thesis
1. Diamond Microfabrication for Applications in Optics and Chemical Sensing
Open this publication in new window or tab >>Diamond Microfabrication for Applications in Optics and Chemical Sensing
2013 (English)Doctoral thesis, comprehensive summary (Other academic)
Abstract [en]

Diamond is a material with many exceptional properties. In this thesis methods for fabrication of microstructures as well as several applications of such structures in optics, microfluidics and electrochemistry are presented.

A method for etching deep and highly precise gratings is described. This method was used to fabricate circularly symmetric half wave plates for use in vector vortex coronagraphs. Such coronagraphs are a very promising approach to the direct imaging of extrasolar planets.

By varying the lateral etch rate of the aluminum mask during diamond etching in an inductively coupled plasma, the sidewall angle of the etched structures could be controlled. This method was used to make smooth sloped sides on a waveguide for coupling light into it. Antireflective structures that drastically reduced the surface reflection in a wavelength band between 10 and 50 µm were also fabricated.

An array of boron doped diamond microelectrodes for electrochemical measurements in a microchannel was fabricated and tested, showing very good stability and reusability. Several hundred hours of use did not adversely affect their performance and no damage to them could be detected by atomic force microscopy or scanning electron microscopy.

Superhydrophobic surfaces in diamond were demonstrated, using both hydrogen and fluorine termination. Hydrogen termination on a flat surface gives contact angles below 90°. To achieve a superhydrophobic surface with this low intrinsic hydrophobicity, structures looking like microscopic nail heads were fabricated. The effect of water pressure on immersed superhydrophobic surfaces was also studied and it was found that the collapse of the superhydrophobic state due to pressure was sometimes reversible as the pressure was lowered.

Finally, a method was tested for functionalizing diamond surfaces using block copolymers of polyethylene oxide and polypropylene oxide to both passivate the surface and to attach synthetic binder molecules. This method was found to give very high signal to noise ratios when detecting C-reactive protein.

Place, publisher, year, edition, pages
Uppsala: Acta Universitatis Upsaliensis, 2013. 65 p.
Series
Digital Comprehensive Summaries of Uppsala Dissertations from the Faculty of Science and Technology, ISSN 1651-6214 ; 1015
Keyword
diamond, microfabrication, microoptics, coronagraph, waveguide, microelectrodes, superhydrophobic
National Category
Manufacturing, Surface and Joining Technology Astronomy, Astrophysics and Cosmology
Research subject
Engineering Science with specialization in Microsystems Technology
Identifiers
urn:nbn:se:uu:diva-192567 (URN)978-91-554-8587-0 (ISBN)
Public defence
2013-03-08, Polacksbacken 2347, Lägerhyddsvägen 2, Uppsala, 10:00 (English)
Opponent
Supervisors
Available from: 2013-02-14 Created: 2013-01-22 Last updated: 2013-04-02

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Fromell, KarinForsberg, PontusKarlsson, MikaelLarsson, KarinNikolajeff, FredrikBaltzer, Lars

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