Detection of mSiglec-E, in solution and expressed on the surface of Chinese hamster ovary cells, using sialic acid functionalised gold nanoparticles

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.3

Copper nanoparticle-induced ovarian injury, follicular atresia, apoptosis, and gene expression alterations in female rats

“…24 The ovarian injury may be associated with alteration of inflammation-related or follicular-atresia-related cytokine expression. 25 In addition, gold NPs can enter Chinese hamster ovary cells through the endocytic pathway, 26 infiltrate rat ovarian granulosa cells and subcellular organelles, and alter in vitro estrogen accumulation. 27 Moreover, mitogen-activated protein kinase cellular signal transduction pathways in mammalian cells can be activated by silicon carbide nanowires.…”

Multivalent Sialosides: A Tool to Explore the Role of Sialic Acids in Biological Processes

“…A similar goal has also driven Haag and co‐workers to synthesize Sia‐conjugated AuNPs (2 and 14 nm) for a sialo‐ligand‐dependent hemaglutination assay with the influenza virus (Figure b) . In yet another development, Russell and co‐workers synthesized sialo‐AuNPs by using direct ligand exchange to detect mSiglec‐E which expressed on CHO cell lines by using a colorimetric method (Figure c) . Overall, direct ligand exchange afforded quick access to the sialo‐AuNPs, which was an efficient platform for the detection of the influenza virus.…”

“…[66] In yet another development, Russell and co-workers synthesized sialo-AuNPs by using direct ligand exchange to detect mSiglec-E which expressed on CHO cell lines by using a colorimetricm ethod (Figure 8c). [67] Overall, direct ligand exchange afforded quick access to the sialo-AuNPs, which was an efficient platform for the detection of the influenzav irus. However,t he major limitation of this strategy was the requirement for al arge quantity of sialosidel igands.…”

Nanotechnology and sialic acid biology

“…Au-NPs functionalized with sialic acid diluted with a PEG forming the sialic acid functionalized gold nanoparticles could detect soluble form of murine Siglec-E (mSiglec-E-Fc fusion protein) on Chinese hamster ovary cells (CHO cells) and find application in detection of Siglec on mammalian cells [87].…”