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Schofield, Claire L., Marin, Maria J. 
ORCID: https://orcid.org/0000-0001-8021-5498, Rejzek, Martin, Crocker, Paul Richard, Field, Robert A. and Russell, David A.
(2016)
Detection of mSiglec-E, in solution and expressed on the surface of Chinese hamster ovary cells, using sialic acid functionalised gold nanoparticles.
The Analyst, 141 (20).
pp. 5799-5809.
ISSN 0003-2654
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Abstract
Sialic acids are widespread in biology, fulfilling a wide range of functions. Their cognate lectin receptors - Siglecs - are equally diverse and widely distributed, with different Siglecs found within distinct populations of cells in the haemopoietic, immune and nervous systems. A convenient way to assay ligand recognition of soluble Siglecs would be useful, as would methods for the concomitant assessment of Siglec distribution on cell surfaces. Here we report the use of gold nanoparticles functionalised with a sialic acid ligand diluted with a polyethylene glycol (PEG) ligand for the plasmonic detection of a soluble form of murine Siglec-E (mSiglec-E-Fc fusion protein) and, importantly, for the specific detection of the same Siglec expressed on the surface of mammalian cells. These sialic acid functionalised nanoparticles are shown to overcome problems such as cellular cis interactions and low Siglec-ligand affinity. The gold nanoparticles were functionalised with various ratios of sialic acid : PEG ligands and the optimum ratio for the detection of murine Siglec-E was established based on the plasmonic detection of the soluble pre-complexed recombinant form of murine Siglec-E (mSiglec-E-Fc fusion protein). The optimum ratio for the detection of the fusion protein was found to be sialic acid : PEG ligands in a 50 : 50 ratio (glyconanoparticles 1). The optimised glyconanoparticles 1 were used to recognise and bind to the murine Siglec-E expressed on the surface of transfected Chinese hamster ovary cells as determined by transmission electron microscopy.
| Item Type: | Article |
|---|---|
| Uncontrolled Keywords: | natural-killer-cells,colorimetric detection,immune system,myleoid cells,carbohydrate microarrays,influenza virus,in vivo,siglecs,binding,glyconanoparticles |
| Faculty \ School: | |
| UEA Research Groups: | Faculty of Science > Research Groups > Chemistry of Materials and Catalysis |
| Depositing User: | Pure Connector |
| Date Deposited: | 31 Oct 2016 17:00 |
| Last Modified: | 22 Oct 2022 01:50 |
| URI: | https://ueaeprints.uea.ac.uk/id/eprint/61186 |
| DOI: | 10.1039/c6an01230b |
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