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  Reversible immobilization of proteins in sensors and solid-state nanopores.

Ananth, A., Genua, M., Aissaoui, N., Diaz, L., Eisele, N. B., Frey, S., et al. (2018). Reversible immobilization of proteins in sensors and solid-state nanopores. Small, 14(18): 1703357. doi:10.1002/smll.201703357.

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Item Permalink: http://hdl.handle.net/21.11116/0000-0000-FFD2-4 Version Permalink: http://hdl.handle.net/21.11116/0000-0003-200A-E
Genre: Journal Article

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2570359.pdf (Publisher version), 3MB
 
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 Creators:
Ananth, A., Author
Genua, M., Author
Aissaoui, N., Author
Diaz, L., Author
Eisele, N. B., Author
Frey, S.1, Author              
Dekker, C., Author
Richter, R. P., Author
Görlich, D.1, Author              
Affiliations:
1Department of Cellular Logistics, MPI for biophysical chemistry, Max Planck Society, ou_578574              

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Free keywords: biosensing; histidine tag; nanopores; selective immobilization; sensors; surface functionalization
 Abstract: The controlled functionalization of surfaces with proteins is crucial for many analytical methods in life science research and biomedical applications. Here, a coating for silica-based surfaces is established which enables stable and selective immobilization of proteins with controlled orientation and tunable surface density. The coating is reusable, retains functionality upon long-term storage in air, and is applicable to surfaces of complex geometry. The protein anchoring method is validated on planar surfaces, and then a method is developed to measure the anchoring process in real time using silicon nitride solid-state nanopores. For surface attachment, polyhistidine tags that are site specifically introduced into recombinant proteins are exploited, and the yeast nucleoporin Nsp1 is used as model protein. Contrary to the commonly used covalent thiol chemistry, the anchoring of proteins via polyhistidine tag is reversible, permitting to take proteins off and replace them by other ones. Such switching in real time in experiments on individual nanopores is monitored using ion conductivity. Finally, it is demonstrated that silica and gold surfaces can be orthogonally functionalized to accommodate polyhistidine-tagged proteins on silica but prevent protein binding to gold, which extends the applicability of this surface functionalization method to even more complex sensor devices.

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Language(s): eng - English
 Dates: 2018-04-032018-05-03
 Publication Status: Published in print
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 Rev. Method: Peer
 Identifiers: DOI: 10.1002/smll.201703357
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Title: Small
Source Genre: Journal
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Pages: 11 Volume / Issue: 14 (18) Sequence Number: 1703357 Start / End Page: - Identifier: -