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Polyelectrolyte-mediated protein adsorption: fluorescent protein binding to individual polyelectrolyte nanospheres

Anikin, K., Röcker, C., Wittemann, A., Wiedenmann, J., Ballauff, M. and Nienhaus, G.U. (2005) Polyelectrolyte-mediated protein adsorption: fluorescent protein binding to individual polyelectrolyte nanospheres Journal of Physical Chemistry B, 109, (12), pp. 5418-5420. (doi:10.1021/jp0506282 S1089-5647(05)00628-0).

Record type: Article

Abstract

We have used confocal fluorescence microscopy with single molecule sensitivity to characterize uptake and release of fluorescent protein (mEosFP) molecules by individual spherical polyelectrolyte brush (SPB) nanoparticles that were immobilized on a glass surface. The SPB particles consisted of a solid core particle of 100 nm diameter onto which long polyelectrolyte chains were affixed. They could be loaded with up to 30 000 mEosFP molecules in a solvent of low ionic strength. The concentration dependence of protein loading can be described with a simple bimolecular binding model, characterized by an equilibrium dissociation coefficient of 0.5 M. Essentially complete release of the bound proteins was observed after increasing the ionic strength by adding 250 mM NaCl to the solvent. Fluorescence emission spectra and time-resolved fluorescence intensity decays were measured on individual, mEosFP-loaded SPB nanoparticles, and also on the dissolved mEosFP before and after adsorption. These results indicate that the mEosFP molecules remained structurally intact in this procedure. Hence, the present investigation demonstrates unambiguously that polyelectrolyte-mediated protein adsorption onto SPB particles presents a viable process for protein immobilization.

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Published date: 2005

Identifiers

Local EPrints ID: 51188
URI: http://eprints.soton.ac.uk/id/eprint/51188
ISSN: 1089-5647
PURE UUID: a9853ac4-f9df-4ff6-a612-45548d690efa
ORCID for J. Wiedenmann: ORCID iD orcid.org/0000-0003-2128-2943

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Date deposited: 08 May 2008
Last modified: 17 Jul 2017 14:49

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Contributors

Author: K. Anikin
Author: C. Röcker
Author: A. Wittemann
Author: J. Wiedenmann ORCID iD
Author: M. Ballauff
Author: G.U. Nienhaus

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