Back to Search
Start Over
Optimization of a non-radioactive high-throughput assay for decarboxylase enzymes.
- Source :
-
Assay and drug development technologies [Assay Drug Dev Technol] 2010 Apr; Vol. 8 (2), pp. 175-85. - Publication Year :
- 2010
-
Abstract
- Herein, we describe the optimization of a linked enzyme assay suitable for high-throughput screening of decarboxylases, a target family whose activity has historically been difficult to quantify. Our approach uses a commercially available bicarbonate detection reagent to measure decarboxylase activity. The assay is performed in a fully enclosed automated screening system under inert nitrogen atmosphere to minimize perturbation by exogenous CO2. Receiver operating characteristic (ROC) analysis following a pilot screen of a small library of approximately 3,600 unique molecules for inhibitors of Trypanosoma brucei ornithine decarboxylase quantitatively demonstrates that the assay has excellent discriminatory power (area under the curve = 0.90 with 95% confidence interval between 0.82 and 0.97).
- Subjects :
- Animals
Bicarbonates analysis
Carboxy-Lyases antagonists & inhibitors
Carboxy-Lyases isolation & purification
Data Interpretation, Statistical
Drug Evaluation, Preclinical methods
Enzyme Assays
Enzyme Inhibitors pharmacology
Malate Dehydrogenase analysis
Ornithine Decarboxylase analysis
Ornithine Decarboxylase metabolism
Ornithine Decarboxylase Inhibitors
Phosphoenolpyruvate Carboxylase analysis
ROC Curve
Trypanosoma brucei brucei enzymology
Carboxy-Lyases analysis
Subjects
Details
- Language :
- English
- ISSN :
- 1557-8127
- Volume :
- 8
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Assay and drug development technologies
- Publication Type :
- Academic Journal
- Accession number :
- 20085486
- Full Text :
- https://doi.org/10.1089/adt.2009.0249