2009
DOI: 10.1002/chem.200802305
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Mapping Receptor Density on Live Cells by Using Fluorescence Correlation Spectroscopy

Abstract: Study of the density, spatial distribution, and molecular interactions of receptors on the cell membrane provides the knowledge required to understand cellular behavior and biological functions, as well as to discover, design, and screen novel therapeutic agents. However, the mapping of receptor distribution and the monitoring of ligand–receptor interactions on live cells in a spatially and temporally ordered manner are challenging tasks. In this paper, we apply fluorescence correlation spectroscopy (FCS) to m… Show more

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Cited by 96 publications
(90 citation statements)
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“…For example, when various concentrations of fluorescein isothiocyanate (FITC)-labeled sgc8 aptamers are incubated with HeLa cells, the different diffusion times of bound versus free aptamers can be used to assess the percentage of bound aptamer, followed by yielding the absolute number of total aptamers inside the confocal volume. A picomolar range dissociation constant (K d = 790 ± 150 pM) of sgc8 aptamer has been measured for HeLa cells, which is in good agreement with the result determined by flow cytometry (K d = 810 pM) [33].…”
Section: K D Determination Of Cell-specific Aptamersmentioning
confidence: 74%
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“…For example, when various concentrations of fluorescein isothiocyanate (FITC)-labeled sgc8 aptamers are incubated with HeLa cells, the different diffusion times of bound versus free aptamers can be used to assess the percentage of bound aptamer, followed by yielding the absolute number of total aptamers inside the confocal volume. A picomolar range dissociation constant (K d = 790 ± 150 pM) of sgc8 aptamer has been measured for HeLa cells, which is in good agreement with the result determined by flow cytometry (K d = 810 pM) [33].…”
Section: K D Determination Of Cell-specific Aptamersmentioning
confidence: 74%
“…Aptamers, which target cellular membrane surface biomarkers, are considered to be better molecular probes for surface biomarkers discovery compared to antibody-based recognition. This is due to the higher binding affinity (low K d ), specificity, and increased tumor penetration associated with the small size of aptamers, as well as the potentially reduced immunogenicity [32,33,36]. The density investigation of cellular receptors provided by the binding events of aptamers to specific receptors can help us to gain insight into membrane receptor characteristics, expression levels, and spatial distribution on the molecular level, as well as receptor clustering and molecular changes in living biological specimens.…”
Section: Significance Of Aptamer Binding Densitymentioning
confidence: 97%
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“…In the present nanotags, Au-Ag NRs were applied as the SERS substrates, and 4-ATP molecules were absorbed onto the NRs to provide SERS signal. The sgc8c aptamers on the nanotags can specifically bind to the PTK-7, which is overexpressed on Hela cell surface with the density of 550±90 receptor μm −2 based on the previous fluorescence correlation spectroscopy (FCS) study [30]. The high density of PTK-7 induced the accumulation of the nanotags into close proximity on the cell surface.…”
Section: Target Cell Detection By the Dual Mode Nanotagsmentioning
confidence: 99%