3 research outputs found
High-Throughput Profiling of Nanoparticle–Protein Interactions by Fluorescamine Labeling
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High-throughput profiling of nanoparticle-protein interactions by fluorescamine labeling.
A rapid, high throughput fluorescence assay was designed to screen interactions between proteins and nanoparticles. The assay employs fluorescamine, a primary-amine specific fluorogenic dye, to label proteins. Because fluorescamine could specifically target the surface amines on proteins, a conformational change of the protein upon interaction with nanoparticles will result in a change in fluorescence. In the present study, the assay was applied to test the interactions between a selection of proteins and nanoparticles made of polystyrene, silica, or iron oxide. The particles were also different in their hydrodynamic diameter, synthesis procedure, or surface modification. Significant labeling differences were detected when the same protein incubated with different particles. Principal component analysis (PCA) on the collected fluorescence profiles revealed clear grouping effects of the particles based on their properties. The results prove that fluorescamine labeling is capable of detecting protein-nanoparticle interactions, and the resulting fluorescence profile is sensitive to differences in nanoparticle's physical properties. The assay can be carried out in a high-throughput manner, and is rapid with low operation cost. Thus, it is well suited for evaluating interactions between a larger number of proteins and nanoparticles. Such assessment can help to improve our understanding on the molecular basis that governs the biological behaviors of nanomaterials. It will also be useful for initial examination of the bioactivity and reproducibility of nanomaterials employed in biomedical fields
High-Throughput Profiling of Nanoparticle–Protein Interactions by Fluorescamine Labeling
A rapid, high throughput fluorescence
assay was designed to screen
interactions between proteins and nanoparticles. The assay employs
fluorescamine, a primary-amine specific fluorogenic dye, to label
proteins. Because fluorescamine could specifically target the surface
amines on proteins, a conformational change of the protein upon interaction
with nanoparticles will result in a change in fluorescence. In the
present study, the assay was applied to test the interactions between
a selection of proteins and nanoparticles made of polystyrene, silica,
or iron oxide. The particles were also different in their hydrodynamic
diameter, synthesis procedure, or surface modification. Significant
labeling differences were detected when the same protein incubated
with different particles. Principal component analysis (PCA) on the
collected fluorescence profiles revealed clear grouping effects of
the particles based on their properties. The results prove that fluorescamine
labeling is capable of detecting protein–nanoparticle interactions,
and the resulting fluorescence profile is sensitive to differences
in nanoparticle’s physical properties. The assay can be carried
out in a high-throughput manner, and is rapid with low operation cost.
Thus, it is well suited for evaluating interactions between a larger
number of proteins and nanoparticles. Such assessment can help to
improve our understanding on the molecular basis that governs the
biological behaviors of nanomaterials. It will also be useful for
initial examination of the bioactivity and reproducibility of nanomaterials
employed in biomedical fields