| dc.contributor.author |
Papaneophytou, CP |
en |
| dc.contributor.author |
Mettou, AK |
en |
| dc.contributor.author |
Rinotas, V |
en |
| dc.contributor.author |
Douni, E |
en |
| dc.contributor.author |
Kontopidis, GA |
en |
| dc.date.accessioned |
2014-06-06T06:52:50Z |
|
| dc.date.available |
2014-06-06T06:52:50Z |
|
| dc.date.issued |
2013 |
en |
| dc.identifier.issn |
19485875 |
en |
| dc.identifier.uri |
http://dx.doi.org/10.1021/ml300380h |
en |
| dc.identifier.uri |
http://62.217.125.90/xmlui/handle/123456789/6203 |
|
| dc.subject |
aqueous solubility |
en |
| dc.subject |
fluorescence binding assay |
en |
| dc.subject |
insoluble ligands |
en |
| dc.subject |
protein stability |
en |
| dc.subject |
solvent selection |
en |
| dc.subject.other |
dimethyl sulfoxide |
en |
| dc.subject.other |
glycerol |
en |
| dc.subject.other |
macrogol 3350 |
en |
| dc.subject.other |
protein inhibitor |
en |
| dc.subject.other |
spd 304 |
en |
| dc.subject.other |
tumor necrosis factor alpha inhibitor |
en |
| dc.subject.other |
unclassified drug |
en |
| dc.subject.other |
article |
en |
| dc.subject.other |
binding assay |
en |
| dc.subject.other |
fluorescence analysis |
en |
| dc.subject.other |
priority journal |
en |
| dc.subject.other |
protein stability |
en |
| dc.title |
Solvent selection for insoluble ligands, a challenge for biological assay development: A TNF-α/SPD304 study |
en |
| heal.type |
journalArticle |
en |
| heal.identifier.primary |
10.1021/ml300380h |
en |
| heal.publicationDate |
2013 |
en |
| heal.abstract |
Many active compounds may be excluded from biological assays due to their low aqueous solubility. In this study, a simple method for the determination of the solubility of compounds containing aromatic rings is proposed. In addition to DMSO, five organic solvents for screening experiments of TNF-α inhibitors were explored. DMSO and PEG3350 were the most suitable for both protein stability and ligand-binding experiments. In addition, glycerol is a promising solvent for the screening of other compounds for which it might provide acceptable solubilization, due to its strong tendency to preserve the protein. Moreover, a fluorescence binding assay was developed using the TNF-α/SPD304 system, and a Kd of 5.36 ± 0.21 μM was determined. The results of this study could be used for the future screening of potential TNF-α inhibitors, while the protocols developed in this work could be applied to other proteins. © 2012 American Chemical Society. |
en |
| heal.journalName |
ACS Medicinal Chemistry Letters |
en |
| dc.identifier.issue |
1 |
en |
| dc.identifier.volume |
4 |
en |
| dc.identifier.doi |
10.1021/ml300380h |
en |
| dc.identifier.spage |
137 |
en |
| dc.identifier.epage |
141 |
en |