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.
Original language | English |
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Pages (from-to) | 5799-5809 |
Number of pages | 11 |
Journal | The Analyst |
Volume | 141 |
Issue number | 20 |
Early online date | 18 Aug 2016 |
DOIs | |
Publication status | Published - 21 Oct 2016 |
Keywords
- natural-killer-cells
- colorimetric detection
- immune system
- myleoid cells
- carbohydrate microarrays
- influenza virus
- in vivo
- Siglecs
- binding
- glyconanoparticles
Profiles
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Maria J. Marin
- School of Chemistry, Pharmacy and Pharmacology - Associate Professor in Analytical Chemistry
- Centre for Photonics and Quantum Science - Member
- Chemistry of Life Processes - Member
- Chemistry of Light and Energy - Member
- Chemistry of Materials and Catalysis - Member
Person: Research Group Member, Academic, Teaching & Research