Cellular uptake of ribonuclease A-functionalised core-shell silica microspheres

G.F Chimonides, J.M. Behrendt, E. Chundoo, C. Bland, A.V. Hine, A. Devitt, D.A. Nagel, A.J. Sutherland

Research output: Contribution to journalArticle

Abstract

Analysis of protein function in a cellular context ideally requires physiologically representative levels of that protein. Thus conventional nucleic acid-based transfection methods are far from ideal owing to the over expression that generally results. Likewise fusions with protein transduction domains can be problematic whilst delivery via liposomes/nanoparticles typically results in endosomal localisation. Recently polymer microspheres have been reported to be highly effective at delivering proteins into cells and thus provide a viable new alternative for protein delivery (protein transduction). Herein we describe the successful delivery of active ribonuclease A into HeLa cells via novel polymer core-silica shell microspheres. Specifically, poly(styrene-co-vinylbenzylisothiouronium chloride) core particles, generated by dispersion polymerisation, were coated with a poly(styrene-co-trimethoxysilylpropyl methacrylate) shell. The resultant core-shell morphology was characterised by transmission electron, scanning electron and fluorescence confocal microscopies, whilst size and surface charge was assessed by dynamic light scattering and zeta-potential measurements, respectively. Subsequently ribonuclease A was coupled to the microspheres using simple carbodiimide chemistry. Gel electrophoresis confirmed and quantified the activity of the immobilised enzyme against purified HeLa RNA. Finally, the polymer-protein particles were evaluated as protein-transduction vectors in vitro to deliver active ribonuclease A to HeLa cells. Cellular uptake of the microspheres was successful and resulted in reduced levels of both intracellular RNA and cell viability.
LanguageEnglish
Pages7307-7315
Number of pages9
JournalJournal of Materials Chemistry B
Volume2
Issue number42
Early online date27 Aug 2014
DOIs
Publication statusPublished - 14 Nov 2014

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Pancreatic Ribonuclease
Microspheres
Silicon Dioxide
Silica
Proteins
Polymers
Styrene
HeLa Cells
RNA
Electrons
Carbodiimides
Immobilized Enzymes
Methacrylates
Ribonucleases
Liposomes
Confocal microscopy
Fluorescence microscopy
Fluorescence Microscopy
Nucleic acids
Confocal Microscopy

Bibliographical note

This article is licensed under a Creative Commons Attribution 3.0 Unported Licence.

Funding: EPSRC Industrial CASE (GFC) grant number EP/D038057/1
Electronic supplementary information: see DOI: 10.1039/c4tb01130a

Cite this

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Cellular uptake of ribonuclease A-functionalised core-shell silica microspheres. / Chimonides, G.F; Behrendt, J.M.; Chundoo, E.; Bland, C.; Hine, A.V.; Devitt, A.; Nagel, D.A.; Sutherland, A.J.

In: Journal of Materials Chemistry B, Vol. 2, No. 42, 14.11.2014, p. 7307-7315.

Research output: Contribution to journalArticle

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AU - Bland, C.

AU - Hine, A.V.

AU - Devitt, A.

AU - Nagel, D.A.

AU - Sutherland, A.J.

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