Identification of Fluorescent Compounds with Non-Specific Binding Property via High Throughput Live Cell Microscopy
SCIE
SCOPUS
- Title
- Identification of Fluorescent Compounds with Non-Specific Binding Property via High Throughput Live Cell Microscopy
- Authors
- Nath, Sangeeta; Spencer, Virginia A.; Han, Ju; Chang, Hang; Zhang, Kai; Fontenay, Gerald V.; Anderson, Charles; Hyman, Joel M.; Nilsen-Hamilton, Marit; Chang, Young-Tae; Parvin, Bahram
- Date Issued
- 2012-01
- Publisher
- PUBLIC LIBRARY SCIENCE
- Abstract
- Introduction: Compounds exhibiting low non-specific intracellular binding or non-stickiness are concomitant with rapid clearing and in high demand for live-cell imaging assays because they allow for intracellular receptor localization with a high signal/noise ratio. The non-stickiness property is particularly important for imaging intracellular receptors due to the equilibria involved. Method: Three mammalian cell lines with diverse genetic backgrounds were used to screen a combinatorial fluorescence library via high throughput live cell microscopy for potential ligands with high in-and out-flux properties. The binding properties of ligands identified from the first screen were subsequently validated on plant root hair. A correlative analysis was then performed between each ligand and its corresponding physiochemical and structural properties. Results: The non-stickiness property of each ligand was quantified as a function of the temporal uptake and retention on a cell-by-cell basis. Our data shows that (i) mammalian systems can serve as a pre-screening tool for complex plant species that are not amenable to high-throughput imaging; (ii) retention and spatial localization of chemical compounds vary within and between each cell line; and (iii) the structural similarities of compounds can infer their non-specific binding properties. Conclusion: We have validated a protocol for identifying chemical compounds with non-specific binding properties that is testable across diverse species. Further analysis reveals an overlap between the non-stickiness property and the structural similarity of compounds. The net result is a more robust screening assay for identifying desirable ligands that can be used to monitor intracellular localization. Several new applications of the screening protocol and results are also presented.
- Keywords
- LINES; LOCALIZATION; KEGG
- URI
- https://oasis.postech.ac.kr/handle/2014.oak/50340
- DOI
- 10.1371/journal.pone.0028802
- ISSN
- 1932-6203
- Article Type
- Article
- Citation
- PLOS ONE, vol. 7, no. 1, 2012-01
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