Engineered self-assembling monolayers for label free detection of influenza nucleoprotein
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Date
2015-06
Journal Title
Journal ISSN
Volume Title
Publisher
Springer Nature
Abstract
Integrating nanotechnology into useable devices requires a combination of bottom up and top down methodology. Often the techniques to measure and control these different components are entirely different, so methods that can analyse the nanoscale component in situ are of increasing importance. Here we describe a strategy that employs a self-assembling monolayer of engineered protein chimeras to display an array of oriented antibodies (IgG) on a microelectronic device for the label free detection of influenza nucleoprotein. The structural and functional properties of the bio-interface were characterised by a range of physical techniques including surface plasmon resonance, quartz-crystal microbalance and neutron reflectometry. This combination of methods reveals a 13.5 nm thick engineered-monolayer that (i) self-assembles on gold surfaces, (ii) captures IgG with high affinity in a defined orientation and (iii) specifically recognises the influenza A nucleoprotein. Furthermore we also show that this non-covalent self-assembled structure can render the dissociation of bound IgG irreversible by chemical crosslinking in situ without affecting the IgG function. The methods can thus describe in detail the transition from soluble engineered molecules with nanometre dimensions to an array that demonstrates the principles of a working influenza sensor. © The Author(s) 2015. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.
Description
Funding from Technology Strategy Board grant (TSB/100565) to Orla Protein Technologies Ltd and Newcastle University for the ‘Virasens’ project and Wellcome Trust Grant Number 093581. The authors thank the Bragg Institute Programme Advisory Committee (proposal P1500), and the ISIS Facility Access Panel (RB121134), for the award of neutron beam time. APLB acknowledges a Discovery Early Career Researcher Award from the Australian Research Council (DE140101788). The authors thank Marie Gillon (ANSTO) and Helen Ridley (Newcastle University) for technical assistance.
Keywords
Influenza, Nanotechnology, Protein engineering, Antibodies, Nucleoproteins, Plasmons, Quartz, Microbalances, Neutron radiography, Quartz
Citation
Le Brun, A. P., Soliakov, A., Shah, D. S. H., Holt, S. A., McGill, A., & Lakey, J. H. (2015). Engineered self-assembling monolayers for label free detection of influenza nucleoprotein. Biomedical Microdevices, 17(3), 49. doi.org/10.1007/s10544-015-9951-z